Agro-Technical and Technology College, Harar (ATTC)

Department of Agro-Ecology

Program: Undergraduate

Seed Science and Technology _ Handout

Instructor: Abiyou D. (MSc.)

March 2022

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 : INTRODUCTION
1.1. Definitions: Science and Technology
Science and Technology are a topic that includes science, technology, and the interactions between science and
technology.

I. Science is a systematic study that builds and organizes knowledge in the form of explanations and predictions about
nature and the universe. Science is defined as a process and a product:

Science as a process:
▪ It is determined by observation, hypothesis, experimentation, measurement and analysis.
▪ Conceptualization of new ideas, from the abstract (theoretical rather than physical) to the particular.
Science as a product:
▪ Systematized and organized form of knowledge based on facts.
▪ Concerned with verifiable or demonstrable concepts, principles, theories and laws.
▪ It is the variety of knowledge, skills, physical resources, and technologies that taken together in relation with
one another.
II. Technology is the collection of techniques, methods or processes used in the production of goods or services.
Technology also defined as a process and a product:
▪ Technology as a process: It is the application of scientific knowledge or science.
▪ Technology as a product: A system of knowledge, skills, techniques and processes.

In broad sense, Science and technology defined as: A system of knowledge, skills, techniques and processes which
enable society to produce, distribute, install, maintain or improve goods and services needed to satisfy human needs.

1.2. Definitions: Seed, Seed Science and Technology

I. What is a seed?
▪ In broad sense, seed is a material which is used for planting or regeneration purpose. A living plant part which when
provided with ideal growing conditions, can produce a fully functional plant.
▪ True seed is a result of pollination and fertilization of the ovule resulting in combination of genetic material thus
causing variation.
▪ Vegetative propagation materials give daughter plants that are identical to the parent plants.
▪ True seed may be defined structurally “It is a fertilized matured ovule, consisting of an embryonic plant, a store
of food in cotyledons &/or endosperm and a protective seed coat.”
▪ However, from the seed technological point of view seed is defined as: Any plant part that is intended for planting
and will include the true seed and vegetative propagation materials such as healthy seedlings, tuber, bulbs,
rhizome, roots, cuttings, all types of grafts and any plant part used for propagation purposes.

Therefore, seed is the most vital and crucial input for crop production, one of the ways to increase the productivity
without adding new cultivation land by planting quality seed. Good quality seeds can contribute to increase 15-20%
yield of agricultural crops. Food security is highly dependent on seed security.

The world is rapidly changing and facing major global challenges:

➢ 820 million people regularly go to bed hungry (98% live in developing countries)
➢ 150 million children under 5 years age go to bed hungry every night.

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In 2050 to feed 9.8 billion people should have to double food production. Climate change will cause multiple impact on
water, health, agriculture and biodiversity. Planet needs bold and solid solutions to turn vision into reality. Currently,
millions of African peasant farmers owning a piece of land. Similarly, Ethiopian farmers also own in average less than
one-hectare land is likely to decline further.

Therefore, to alleviate the coming disaster science-based solutions is important to increase food production in the world,
especially in the developing countries. Seed is one of the key inputs to improve crop production and productivity.
Increasing the quality of seeds can increase the yield potential of the crop by significant folds. Quality seed is one of the
most economical and efficient inputs to agricultural development. As a key agricultural input, seeds play a fundamental
role in meeting the triple challenge of improving food security and nutrition, supporting the livelihoods of farmers and
rural communities, and contributing to sustainable resource use and climate change adaptation and mitigation
(Eniviromental sustainability). Generation and transfer of improved technologies are critical prerequisites for
agricultural development particularly for an agrarian based economy such as of Ethiopian.

Seed and grain

We often encounter two terms seed and grain. The biological process by which seeds and grains are formed is basically
the same, but the two are quite different with respect to functions and objectives of production. Seed is meant to be used
for producing good crops whereas grain is meant for food, feed or raw material. Therefore, both commodities have to
be handled differently.

Seed and grain production follow similar operations but different strategies. Apart from good agronomic management
of the crop, seed production differs from grain production on the following key issues: land requirement, isolation,
roguing, prevention of contamination and limitations of generations. Another difference is that seed crops must meet
specific quality standards prescribed by the national seed regulations. The technical, administrative and legislative
control by the certification agency provides guidelines that have to be followed to produce good quality seed that meets
the standards.

The differences between seed and grain are given below:

Seed Grain
1 It should be a viable. Embryo is important. It is used for Need not be a viable. Endosperm is important.
planting purpose. It is used for consumption
2 It should have maximum genetic & physical purity Not so
3 Should satisfy minimum seed certification standards No such requirements
4 Treated with pesticide /fungicide to protect seed against Not treated with any chemicals, since used for
storage pests and fungi consumption
5 Respiration rate and other physiological and biological No such specifications
processes should be kept at low level during storage
6 Production is technically organized Not so
7 It should satisfy all the seed quality attributes No need
8 Seed can be used as grain, if it is not treated with poisonous Grain never can be used as seed.
chemicals.

In modern agriculture, seed is a vehicle to deliver almost all agriculture-based technological innovations to farmers so
that they can exploit the genetic potential of new varieties. The availability, access and use of seed of adaptable modern

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varieties is, therefore, determinant to the efficiency and productivity of other packages (irrigation, fertilizers, pesticides)
in increasing crop production to enhance food security and alleviating rural poverty in developing countries.

For seed to play a catalytic role, it should reach farmers in a good quality state, i.e., high genetic purity and identity, as
well as high physical, physiological and health quality. Therefore, the best production techniques need to be followed
to produce good quality seed.

The seed plants are often divided arbitrarily into two groups: the gymnosperms and the angiosperms. The basis for
this distinction is that angiosperms produce flowers, while the gymnosperms do not.
A. Angiosperms: are the flowering plants. This are the largest and most diverse group within the kingdom plantae. With
around 300,000 species, they represent approximately 80 percent of all the known green plants that exist now. The seeds
of this group of plants found inside the fruit. Angiosperms are subdivided into two identifiable groups monocotyledons
and dicotyledons.
Difference between Monocotyledons and Dicotyledons

B. Gymnosperms: are non-flowering plants. Gymnosperms are a smaller, more ancient group, and it consists of plants
that produce “naked seeds” (seeds that are not protected by a fruit). Gymnosperm seeds are usually formed in unisexual
cones, known as cone/strobili, and the plants lack fruits and flowers. There are more than 1,000 species of gymnosperms
still found on Earth. This group include all of the conifers: cedar, redwood, juniper, cypress, fir and pine.

II. Seed Science: the study of the structure and development of seeds from the moment of fertilization of the egg cell
on the maternal plant until formation of a new plant from the seed. The science is closely connected with botany,
biochemistry, genetics, and other biological sciences.

III. Seed Technology

Seed Technology defined as “discipline of studies having to do with seed production, maintenance, quality and
preservation” (Cowan, 1973). The role of seed technology is to protect the biological entity of seed.

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Seed technology as “the methods through which the genetic and physical characteristics of seeds could be improved. It
involves activities such as variety development, evaluation and release, seed production, processing, storage and
certification” (Feistritzer, 1975).

Generally, Seed technology includes the development of superior crop plant varieties, their evaluation and release, seed
production, processing, seed storage, seed testing, seed quality control, seed certification, seed marketing, distribution
and research on seed.

Seed technology is an interdisciplinary science. The relation of seed technology with other disciplines: Plant breeding
and genetics; Agronomy; Horticulture; Plant pathology; Entomology; Taxonomy; Seed physiology and ecology;
Economics; Seed processing and storage/Engineering

Role of Seed Technology

Feistritzer (1975) outlined the following as roles of improved seed.
▪ A carrier of new technologies.
▪ A basic tool to secure food supply.
▪ The principal means to secure crop yields in less favorable production areas.
▪ A medium and rapid rehabilitation of agriculture in cases of natural disaster.

Goals of Seed Technology

The major goal of seed technology is to increase agricultural production through spread of good quality seeds of high
yielding varieties (HYV).
Therefore, the broad objectives of seed technology are given below

A. To assure rapid seed multiplication of desirable varieties.

B. Timely supply of seeds, i.e., well before the sowing season.
C. Supply high quality of seeds: means seeds of high yielding varieties, varieties with resistance to diseases and pests.
D. Supply of seeds at reasonable prices.

Benefits of using quality seeds

✓ They are genetically pure (true to type).
✓ The good quality seed has high return per unit area as the genetic potentiality of the crop can be fully exploited.
✓ Less infestation of land with weed seed/other crop seeds.
✓ Faster growth rate, and greater resistance to stress and diseases
✓ A higher percentage and uniform emergence of seedling.
✓ A vigorous seedling establishment, free from pests and disease.
✓ They can be adopted themselves for extreme climatic condition and cropping system of the location.
✓ The quality seed respond well to the applied fertilizers and nutrients.
✓ Uniform in plant population and maturity.
✓ Handling in post-harvest operation will be easy.
✓ High produce value and their marketability.

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 : FLOWERING PROCESSES, SEED FORMATION, DEVELOPMENT AND
MATURATION IN PLANTS
2.1. Flowering Processes in Plants

Plant growth originates within the buds in regions known as meristems. In the meristems, cell division and elongation
occur, and these processes produce tissues that soon develop into specific plant parts. Vegetative meristems give rise
to parts such as stems, leaves, and roots, while reproductive meristems give rise to floral organs that ultimately produce
fruits and seeds. Within every meristem are minute primordia that resemble twisted outgrowths or ribbed inverted cones.
Although not distinguishable to the naked eye, the configurations of the primordia become visible when the bud scales
are removed and examined under magnification. As growth proceeds, the configurations enlarge and differentiate into
recognizable plant organs. The following two changes are important for the formation of reproductive meristem:

2.1.1. Floral induction

Floral induction is physiological changes in response to external stimuli (light quality, daylength, etc.) that occur in
vegetative meristems of plant and subsequently allow them to become reproductive meristems and undergo floral
initiation. The ability to support reproductive processes requires tremendous energy. Often, many crops do not begin to
form flowers, and eventually seeds, until after substantial vegetative growth has occurred. In some cases, as with most
annuals, this is at the end of the life cycle. In other cases, the plant may not become reproductive until after several
growing seasons as with many fruit trees. During this phase, in which the plant is unable to form flowers because it does
not possess sufficient vegetative structure, it is said to be in a juvenile state. However, at some point, enough vegetative
growth occurs and plants reach sexual maturity and are able to flower. After that stage, certain external (or internal)
stimuli can trigger floral induction, a physiological change that permits the development of reproductive primordia. This
change may lead actual flowering by several days, weeks, or even months.

The following external (or internal) stimuli can activate floral induction
A. Temperature Stimuli: - For floral induction to occur, many plants require exposure to low temperatures. This
process has been called vernalization. In its narrowest sense, vernalization means the promotion of flowering in some
winter cereals by cold treatment of the moistened or germinating seeds. In a broader sense, vernalization means the
induction of flowering in any winter annual, biennial, or even perennial species through exposure to low temperatures.
For example, rye, a winter annual, and perennial ryegrass both must undergo prolonged exposure to low temperatures
before they can produce flowers. Sugar beets and carrots are examples of biennial species that grow vegetatively in the
first year, after which they are vernalized by exposure to winter temperatures. The optimum temperature for
vernalization is between 1°C and 7°C. These temperatures must be experienced by the vegetative meristems for periods
of between 10 and 100 days before a reproductive meristem is initiated when the crop is returned to warm temperatures.
In tomato, floral induction is accomplished by repeated exposure to low night temperatures, separated by periods of
higher temperature. This phenomenon occurs in many plants and has been called thermoperiodism.

B. Day-Length Stimuli: - In many species, floral induction occurs in response to day length, or photoperiod. Thus,
plant species have been categorized according to their day-length requirements as short-day, long-day, intermediate-
day, or day-neutral; however, it is really the length of the night, or dark period, that is the critical factor that influences
flowering. The photoperiod requirements for flowering may be qualitative or quantitative. Some short-day plants such
as some soybean varieties are unable to flower except under short-day treatments; in other short-day species, such as
sunflower, flowering is hastened by the appropriate short-day conditions, although it can eventually occur without them.

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C. Chemical Stimuli: - Certain natural and synthetic chemical substances can cause floral induction. Some are auxin
like compounds for example, indoleacetic acid, or the common herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D). At
certain concentrations, gibberellic acid may also cause floral induction. It promotes flowering of long-day plants held
under short-day conditions; however, it inhibits flowering of short-day plants under the same conditions.

D. Nutritional Status: - In floral induction, the nutritional status of a plant is also important, since construction of the
flowering parts is dependent on food availability and translocation. The carbon-to-nitrogen (C: N) ratio is particularly
influential; in species, such as holly, that bear male and female flowers on separate plants, a high nitrogen-to-carbon
ratio favors pistillate rather than staminate flowers. In tomatoes, carbohydrate deficiencies cause microspore
degeneration, leading to pollen sterility; however, a nitrogen deficiency has no such effect.

2.1.2. Floral initiation

Floral initiation is morphological changes in the development of a reproductive meristem from a vegetative meristem.
Following floral induction, which may be triggered by external stimuli, floral initiation is the morphological expression
of the induced state and usually occurs more or less deeply within the meristems of a plant.

In monocotyledonous species, or flowering plants in which a single embryonic seed leaf appears at germination, floral
initiation begins in specialized meristems called dermatogen, which also give rise to the epidermis.
In dicotyledonous species, or flowering plants in which a pair of embryonic seed leaves appear at germination, floral
initiation occurs in the lateral / axillary bud and terminal / apical bud.

Early in their development, reproductive meristems are similar to vegetative meristems, appearing as twisted or ribbed
configurations on an inverted cone or pedestal. As development proceeds, these configurations develop into recognizable
flower parts.

Floral biology
Flower is a reproductive organ bearing pistil/carpel, stamen and usually sepals and petals. Male part of a flower
is androecium consisting of anthers and filament. Female part is gynoecium consisting of ovary, style and stigma. The
flower is said to be perfect, when they contain both male and female parts. A flower with both functional male and
female is called as bisexual. Sometimes in bisexual flower male or female mature slightly at different times. This
nature is called dichogamy which favors cross-pollination, on the contrary prevents self-pollination. If male matures
first it is called as protandry, if female matures first, it is called protogyny.

Figure: Typical structures of a perfect flower

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Imperfect flowers have either male (staminate flower) or female (pistillate flower) part. Such flowers are called as
unisexual flowers. When male and female organs occur in separate flowers on the same plant known as monoecious.
e.g., maize, if they occur in different plants known as dioecious. e.g., papaya.

Development of female and male gametophyte

A. Development of female gametophyte

Seeds of angiosperm originate from meristematic tissue of the ovary wall called Ovule primordia. Within the nucellus,
one cell known as Archesporial cell (2n) develops a special characteristic that distinguishes it from the adjacent cells. It
develops larger than the surrounding cells, having a large nucleus and denser cytoplasm called Megaspore mother cell
(MMC). MMC undergoes a meiotic division, giving rise to four megaspores, each containing a haploid
chromosome set. Among the four cells one functional megaspore survive to give rise to an embryo sac (mature female
gametophyte). While other three megaspore aborts/degenerate. The process of development of functional megaspore
from megaspore mother cell is known as megasporogenesis. The nucleus within the functional megaspore undergoes
three successive mitotic divisions to form eight nuclei. Embryo sac consists of 7 cells but it contains eight nuclei. Eight
nuclei are arranged as three antipodal cells at the chalaza end, one central cell with two polar nuclei, one egg cell
and two synergids cell at the micropylar end. The process of development of eight nuclei in the embryo sac from the
functional megaspore is known as magagametogenesis.

B. Development of Male gametophyte

The microspore mother cell present in anther tissue undergoes a meiotic division to form 4 haploid functional
microspores or pollen grains. This process is known as microsporogenesis. Each of the four microspores is normally
functional and undergoes two divisions, known as microgametogenesis, giving rise to a microgametophyte, or
immature pollen grain. The nucleus of a pollen grain divides mitotically to form a vegetative nucleus (tube nucleus)
and a generative nucleus. The generative nucleus gets surrounded by cytoplasm to become generative cell. At this
stage, pollen grain is two celled a large vegetative cell and a small lenticular generative cell. The pollen grain may be
discharged from the anther at this stage (two-celled stage). However, in some plants, generative cell divides further to
give rise to two sperm nuclei before the pollen grains are shed. These pollen grains are therefore, three-celled at the
time of shedding. In most of the plants, three-celled condition is formed after pollen grain has been shed. The liberated
pollen grain germinates on the stigma and produces a pollen tube. The pollen tube is covered by intine. It secretes
pectinases and other hydrolytic enzymes to create a passage for it in the style. The pollen tube absorbs nourishment from
the cells of the style for its growth.

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2.2. Seed Formation

A true seed is defined as a fertilized mature ovule consisting of embryo, stored food material and protective coats. The
important events involved in seed formation includes
I. Pollination
II. Fertilization
III. Development of the fertilized ovule by
▪ Cell division
▪ Accumulation of reserve food material
▪ Loss of moisture content (Dehydration)

I. Pollination

Pollination is the act of transferring pollen grains from the male anther of a flower to the female stigma. Seeds usually
developed from the fertilized flower for which pollination is prerequisite. For successful fertilization viable pollen and
receptive stigma are two prerequisites. The mature anthers dehisce and release pollen grains (haploid microspores).
When pollen grains are transferred from an anther to the stigma of the same flower the process is called self-
pollination or autogamy (pollination of the ovules of a flower by its own pollen). If they are transferred to the stigma
of another flower, cross-pollination or allogamy (opposed to autogamy).

Generally, self-pollination means the transfer of pollen from the anther to the stigma of the same flower, or another
flower on the same plant. Self-pollination occurs in those plants where bisexual flowers achieve anther dehiscence and
stigma receptivity simultaneously. The majority of angiosperms bear chasmogamous flowers i.e., flowers do not open
before pollination. In some plants, flowers do not open at all such flowers is called cleistogamous, and this is the most
efficient floral adaptation for promoting self-pollination.

Cross-pollination is ensured in plants which bear unisexual flowers. In bisexual flowers also self-pollination may be
prevented by different factors:

A. Self-sterility: inability to produce viable pollen. e.g., Sunflower

B. Dichogamy: maturation of male and female organs at different times. e.g., Pearl Millet
C. Herkogamy: the stamens and the stigma are separated by space so that the stigma cannot get fertilized by the
anther. Different relative positionings of the stigma and the anthers.

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D. Heterostyly: In such flowers the stigma and anthers grow at different heights which does not favor self-pollination.
The condition of having unequal styles e.g., Brassica
E. Self-incompatibility: Such flowers unable to be fertilized by its own pollen. e.g., Cole crops/Brassica

The important self-pollinated crops are wheat, rice, barely, mung bean and cowpea and cross pollinated are maize, rye,
forage legumes and vegetables like carrot, cauliflower and onion. There is yet another category of crops called often
cross-pollinated crops such as cotton and pigeon pea where there may be 10-40% cross pollination.

II. Fertilization
In plants, fertilization is a process of sexual reproduction, which occurs after pollination and germination. Fertilization
can be defined as the fusion of the male gametes (pollen) with the female gametes (ovum) to form a diploid zygote.
Ovules are developed in the female gametophyte and pollen in the male gametophyte. Fertilization always takes place
in female gametophyte; therefore, pollen must have transferred from male to female by pollen vector which may
be abiotic including wind (anemophily) and water (hydrophily) or biotic including insects (entomophily) and bats
(chiropterophily).
Pollen after landing on the stigma, the pollen grain germinates and pollen tube grows through the style. The surface of
the stigma secretes substances, which may provide optimum conditions for pollen germination. The pollen tubes
traversing the style, pectinase which dissolves intercellular substances of the style tissue. After traversing the style, the
pollen tube enters embryo sac of the ovule. The embryo sac consists of 7 cells. The end near the micropyle has the egg
apparatus, which consists of egg cell and 2 synergids. There are 2 polar nuclei in the center and the chalaza end has 3
antipodal cells.

In angiosperms, fertilization involves the participation of 2 male nuclei (double fertilization). One fuses with the egg
nucleus to form the diploid zygote and the other with 2 polar nuclei to produce a triploid nucleus, which is the primary
endosperm nucleus. This process is known as double fertilization.

Figure: Shows the various parts of a fully formed ovule

Development of seed without fertilization i.e., the seed formation occurs without sexual fusion, the process is known as
Apomixis. This process takes place when the megaspore mother cell does not undergo meiosis or a cell from the nucellus
develops into the embryo. This occurs by several mechanisms; however, all apomictic seed have genetic material only
from the female plant. Apomixis may or may not require pollination and pollen tube germination to initiate seed
formation, however sexual union never occurs.

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Parthenocarpy is the production of fruits from unfertilized ovules in plants. The fruits produced by parthenocarpy do
not contain seeds. For example, seedless watermelon and orange. Parthenogenesis is a type of reproduction in which
the ovum develops in to an individual without fertilization.

2.3. Seed Development

After fertilization, development of fertilized ovule into a mature seed involves several different stages. Seed formation
begins within the minute embryo sac with certain expectations, which is about the same in shape, size, and
arrangement. In spite of initial similarities, the seed develops according to the genetic specification for each species,
which are coded in the nucleus (chromosomes) of each cell.

• The integument of the ovule becomes the seed coat of the mature seed.
• Normally the nucellus is absorbed and is absent. The nucellus may persists in some genera such as Papaya, Pepper,
Beet root, etc. as a thin layer called Perisperm, lying inside the seed coat and supplies food material to the embryo.
• The endosperm serves as a principal nutritive support for the embryo of many species (especially monocotyledons)
during both seed development and germination. The endosperm normally grows more rapidly than embryo.
• In monocotyledons, endosperms usually reach the maximum morphological development at physiological maturity
and persists to comprise a major part of the seed.
• In dicotyledonous spp., the endosperm may not develop or may be used up by the developing embryo and contain
none or a small part of mature seed.

A. Embryo development
The first few cell divisions from the zygote forms the Proembryo. Although the mature embryo of monocotyledons
and dicotyledons appears considerably different, their pattern of embryogeny is similar. The Proembryo is divided into
Suspensor and Embryo proper. The suspensor forms into a chain of cells, pushing the embryo proper into the center
of the ovule thus making it in contact with the available food supply. The proembryo may vary greatly in size and shape.

Dicot embryogenesis at proembryo stage

Embryogenesis goes through proembryo, globular, heart, torpedo, and cotyledon stages. Following fertilization of the
egg and sperm nuclei, a proembryo is formed by a transverse cell division to form an apical and basal cell. The basal
cell forms the suspensor, while the apical cell forms the embryo. The suspensor in dicots is usually a column of single
or multiple cells. The suspensor functions to push the proembryo into the embryo sac cavity and to absorb and
transmit nutrients to the proembryo.

Figure: Shows Development of Seed

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Monocot embryogenesis at proembryo stage
Monocots have a more complex embryo structure in the mature seed compared to dicots, but early embryo development
is similar to dicots. The stages of embryogenesis in monocots include the proembryo, globular, scutellar and coleoptilar
stages. Following fertilization, the first cell division is asymmetrical and leads to an apical and basal cell in corn (Zea
mays). Apical cell divides into 2 by a transverse division. The apical cell divides more rapidly than the basal cell and
will eventually be the embryo (In monocotyledons, the basal cell remains undivided). The embryo development in
grasses is different from that of other monocotyledons. The single cotyledon is reduced to absorptive scutellum and
additional structures like coleoptile and coleorhiza are formed.

▪ Apical cell: The small apical cell is on the top and contains most of the cytoplasm, the aqueous substance found within
cells, from the original zygote. It gives rise to the hypocotyl, shoot apical meristem, and cotyledons.
▪ Basal cell: The large basal cell is on the bottom and consists of a large vacuole and the suspensor.

C. Endosperm Development

Endosperm is the nutritive tissue that surrounds and nourishes the embryo in the seeds of angiosperms (flowering plants).
In some seeds the endosperm is completely absorbed at maturity (e.g., pea and bean), and the fleshy food-storing
cotyledons nourish the embryo as it germinates.

There are 3 types of endosperm development:

▪ Nuclear endosperm - where the endosperm nucleus undergoes several divisions prior to cell wall formation. It is the
most common type of endosperm in most of the plants. Commonly referred to as liquid endosperm (Coconut water).
e.g., wheat, apple, squash.
▪ Cellular endosperm - in which there is no free nuclear phase each nuclear division is accompanied by cell wall
formation.
▪ Helobial endosperm - Intermediate between the nuclear and cellular endosperm types in which development is
characterized by free nuclear division as well as cell wall formation in some areas. During the course of seed
development, reserve food materials are accumulated in the endosperm from the adjacent tissues. It is the least
common type of endosperm.

In endospermic dicot seeds, endosperms are retained as a permanent storage tissue. In non-endospermic dicot seeds,
endosperm reserves are depleted and blocked by the developing embryo (temporary storage). The reserves are then
reorganized in the cotyledons, which in turn act as the source of stored reserved food for embryo after germination. A
part of the endosperm is depleted in cereals during embryo maturation and this lies as a layer between the starchy
endosperm and scutellum.

C. Seed-coat Development

Integuments of the ovule undergo marked reorganization and histological changes during maturation to form seed coats.
In bitegmic ovules (which have 2 integuments), the seed coat may be derived from the outer and inner integuments e.g.,
angiosperms. Unitegmic ovules having only one integument, e.g., gymnosperms. The seed coat may be derived from
the outer integument only; the inner integument may disintegrate. Ovules that lack integuments are known as ategmic
ovules as in some of the parasites

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2.4. Seed Maturation

During seed maturation:

▪ The cell cycle ceases
▪ Water content decreases
▪ Storage products are synthesized
▪ Free ABA accumulates mainly to prevent viviparous, and
▪ Primary dormancy is established

Wheat and soybean representing monocots and dicots may illustrate the changes in the pattern of accumulation of reserve
materials at different stages of seed maturation.

In wheat, the dry weight of the seed increases rapidly in about 35 days after anthesis. The water content of the grain is
maximum between 14 and 21 days after anthesis, and then it declines rapidly. The amounts of reducing sugar and sucrose
are high between 7 and 14 days and decline rapidly thereafter due to conversion to starch. Since in wheat, starch is the
major reserve material of the seed, the pattern of starch accumulation is similar to that of dry matter accumulation.

The speed of germination is faster in wheat varieties that begin to lose water early during seed development. The seed
is said to have physiologically matured only when it attains maximum dry weight, germinability and vigour. Normally
the seed is harvested at field maturity, a stage when the moisture content is reduced to about 6-10 % in wheat. Field
maturity is a crop specific character.

A soybean seed attains maximum dry weight between 48 and 54 days after flowering. Oil accumulation is less during
12-18 days after fertilization; maximum oil accumulates between 24 and 42 days after flowering, after which the rate
decreases. The protein content in the seed is maximum during 12-18 days after fertilization and decreases subsequently.
The initial high percentage of protein may be due to the high content of non-protein nitrogen, which decreases with seed
age. Oil accumulation picks up only after protein accumulation completes in the seed.

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 : SEED DORMANCY
3.1. Definition of Seed Dormancy

The ability of seeds to delay their germination until the time and place are right. It is an important survival mechanism
in plants. Seed dormancy may be a complex and puzzling challenge to the seed analyst and the seed researcher, but it is
the method through which plants are able to survive and adapt to their environment. It is common observations that
seeds of many plants’ species do not germinate when freshly harvested even under favorable conditions. They need a
period of rest/ storage before they become capable of germination.

Seed dormancy is defined as a seed failing to germinate under environmental conditions optimal for germination,
normally when the environment is at a suitable temperature with proper soil moisture, light and gases. This kind of
dormancy usually is of most interest to seed technologists, biologists and ecologists.

Seed dormancy is a genetically inherited trait whose intensity is modified by the environment during seed development.
Plants with a long history of domestication generally show less seed dormancy than wild or more recently domesticated
species. When domesticated species exhibit dormancy, they become a problem to seed producers, their customers, and
seed analysts. However, a degree of dormancy in certain crops (e.g., winter cereals) is desirable since it prevents pre-
harvest sprouting and helps maintain seed quality. However, dormancy may cause seeds of numerous species to remain
ungerminated in the soil from a few weeks to several years.

A common misconception of seed dormancy is that it is just a resting state in the absence of suitable germination
conditions. This state is often called quiescence. However, true dormancy is defined as a state in which seeds are
prevented from germinating even under environmental conditions normally favorable for germination. Seed dormancy
in all its forms consequently tends to prevent seeds from germinating in the wrong place at the wrong time.

3.2. Biological Significance of Seed Dormancy

▪ Storage life of seeds is prolonged: it is a survival mechanism prevents germination of all seeds at same time
(staggering germination). In some cases of dormancy one year’s seeds do not germinate the same year, this improves
species survival.
▪ Seed embryo survives during adverse conditions of weather, which are not favorable for growth.
▪ Creation of a seed bank
▪ Prevents the in-situ germination i.e., vivipary

3.3. Seed Dormancy Classification

A number of classification schemes have been proposed to describe seed dormancy.

There are five classes of dormancy based on their mode of action:
▪ Physical dormancy (PY),
▪ Physiological dormancy (PD),
▪ Morphological dormancy (MP),
▪ Morphophysiological dormancy (MPD), and
▪ Combinational dormancy (PY +PD), including both primary and secondary dormancy, occur in seeds.

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An additional classification is based on the timing of the occurrence dormancy: primary dormancy and secondary
dormancy.
▪ Primary (or innate or inherent or true) dormancy (as a result of chemicals/ anatomical features of seed): refers to
the type of dormancy that occurs prior to dispersal as part of the seed developmental program. The majority of mature
seeds show innate (or inherent or primary) dormancy, a condition necessary to prevent germination on the mother
plant before the seed is shed. Innate seed dormancy in most species tends to be gradually lost with time (except in
some tree species), often at a rate depending on temperature and moisture content.

▪ Secondary (or enforced or imposed or induced) dormancy or quiescence state (as a result of unfavorable
environmental conditions): refers to the acquisition of dormancy in a mature seed after imbibition as a result of the
lack of proper conditions for germination. Secondary (or induced) dormancy, in many respects similar to innate
dormancy, may subsequently be induced by conditions which signal that the current environment is inappropriate for
germination. e.g., high temperatures or anaerobic conditions.

Seed dormancy is also divided into three major categories based on parts of the seed that produce dormancy: exogenous,
endogenous and combinational dormancy.

I. Exogenous Dormancy
Dormancy is due to some features of the seed located outside the embryo. This type of dormancy is generally related to
physical properties of the seed coat. Exogenous dormancy is a condition in which the essential germination factors (e.g.,
water, light, & temp.) are not available to the seed & thus it fails to germinate.

A. Physical dormancy/Hard seed coat

✓ Impermeability of seed coat to water: due to seed coat structure, which is hard enough to restrict the entry of
moisture into the seeds, thereby preventing seed germination. e.g., Fabaceae or Leguminosae family
✓ Impermeability of seed coat to gases: is related to the insufficient intake of oxygen by seeds due to impermeability
of seed structure enclosing embryo. e.g., Gramineae, fruit crops & forest trees
B. Mechanical dormancy (Due to resistances of seed coat): preventing expansion of embryo is checked due to
extremely hard seed/fruit structure such as seed coat, endosperm and pericarp etc., e.g., Acacia species
C. Chemical dormancy (Due to presence of growth inhibitors in seed coat or endosperm): biochemical substances
present in seed coat or endosperm block the germination of embryo. e.g., Abscisic acid
e.g., Barley: Aflatoxin; Squash: -Dichlobenil; Tomato: Feruline and Caffeine acid and All species: Coumarin

II. Endogenous dormancy

The reason for dormancy is inhibitors present within the embryo. This dormancy is the most predominant dormancy
found in seeds and is due to the inherent properties of the seed. For example, the seed may possess an excess of inhibitors
that must be removed or reduced prior to germination. Environmental conditions during seed development and
maturation influence the duration of endogenous dormancy. Among these factors are day length, moisture status,
position of the seed in the fruit or inflorescence, age of the mother plant, and temperature during seed maturation.

A. Morphological dormancy (Due to under developed embryo): due to an incomplete development of the embryo
but it is differentiated. In such cases, germination does not occur until the embryos develop to their normal size (growth).
In some species, the embryo is just a mass of cells when seeds are dispersed; it is not differentiated. Before germination
can take place, both differentiation and growth of the embryo have to occur. e.g., Apple

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B. Physiological dormancy (Due to presence of growth inhibitors within the embryo): Dormancy arises from
metabolic blocks produced by biochemical substances called inhibitors present within the embryo like ABA, coumarins,
phenols etc., In such cases germination can begin only when these inhibitors are leached out of the embryo.

C. Combined dormancy (morpho-physiological dormancy)

III. Combinational Dormancy (combination of exogenous (physical) and endogenous (physiological) dormancy)
Combination of two or more types of dormancies is known as double dormancy. It can be morpho-physiological i.e.,
combination of under developed embryo and physiological dormancy or exo-endodormancy i.e., combination of
exogenous and endogenous dormancy conditions i.e., hard seed coat (physical plus intermediate physiological
dormancy).

Seed dormancy is believed to be regulated by a balance of endogenous and other growth inhibitors and promoters. The
level of these substances is controlled by certain environmental stimuli & these substances could be:

➢ Germination inhibitors: Cyanide, Phenolics, Abscisic acid (ABA: inhibits the transcription of genes as a result
proper enzyme for seed germination are not produced).
➢ Germination Promoters: Gibberellic acid and Cytokinin. (Gibberellin induces seed germination by increasing the
concentration of Gibberellin than ABA within seed this makes the seed active for germination by removing
dormancy).

3.4. Factors that Control Dormancy Induction

I. Primary dormancy is commonly associated with the transient increase in ABA content during seed development. In
most of the species studied, ABA levels increase during the first half of seed development and decline during late
maturation alongside with the decline in seed water content. ABA has been detected in all seed and fruit tissues examined
and has been related to a number of developmental processes, including synthesis of storage proteins and late
embryogenesis abundant proteins, suppression of precocious germination, and induction of desiccation tolerance. ABA-
deficient mutants of Arabidopsis and tomato have demonstrated that defects in ABA synthesis during seed development
result in the formation of nondormant seeds. ABA content and germination (%) in developing seeds are inversely
correlated.

II. The term secondary dormancy is used for the type of dormancy that is imposed after seeds have lost primary
dormancy. Secondary dormancy is caused by conditions after the seed has been dispersed and occurs in some seeds
when nondormant seed is exposed to conditions that are not favorable to germination, very often high temperatures. The
mechanisms of secondary dormancy are not yet fully understood, but might involve the loss of sensitivity in receptors
in the plasma membrane.

Sometimes nondormant seeds encounter conditions that subsequently cause them to become dormant. This may be
caused by exposure of the seed to conditions that favor germination in all respects except one. Induction of secondary
dormancy was possible one and one-half months after seeds reached physiological maturity, although the persistence of
dormancy obtained in this way decreased almost continuously as the time between physiological maturity and treatments
increased. Secondary dormancy as being either thermo (temperature), photo (light), or skoto (darkness) imposed, though
other causes such as imposition by excess or adverse amounts of water, chemicals, and gases might also be involved.

Two suggestions have been made to explain the mechanism of secondary dormancy:
A. The imposition of a block at crucial points in the metabolic sequence leading to germination, or
B. An unfavorable balance of growth-promoting versus growth-inhibiting substances.

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3.5. Methods to Overcome Seed Dormancy

I. Natural breaking of seed dormancy: in nature dormancy terminates when embryo gets suitable environment such
as adequate moisture, aeration and temperature. The impermeable seed coat present in many species became permeable
due to the rupturing of softening action of natural agents like microorganism, high or low temperature fluctuations,
natural soil acidity, humidity fiber and abrasion due to wind or digestive tracts of birds and animals which feed on these
seeds. e.g., Rhizoctonia damages seed coat

II. Artificial overcoming of seed dormancy: the various treatments for overcoming dormancy may be divided into
the following three groups

A. Seed coat treatments

These treatments aim at making hard seed coat permeable to water or gases either cracking or softening them. The
process is usually referred as scarification. These treatments are either physical or chemical in nature.

Scarification
▪ Acid scarification: treating seeds with concentrated acids like sulphuric acid, Hydrochloric acid etc.,
▪ Thermal scarification: the seeds are treated with different temperatures and gases
▪ Mechanical scarification: The seed coat is damaged using mechanical means. Rubbing seeds by sand paper or by
using mechanical scarifier, piercing by needle and removing of entire seed coat.

B. Embryo treatments
▪ Cold stratification: the incubation of seeds at a suitable low temperature (usually 0- 5C) over a moist substratum
before transferring them to a temperature optimum for germination. e.g., Mustard and rape seeds
▪ High temperature treatment: in some species, incubation at 40-50C for few hours to 1-5 days may be effective in
overcoming dormancy. e.g., Rice seeds more than 15% seed moisture treated in hot water of 40C for 4-5 hours.
Thermo dormancy is seed sensitivity to heat or cold.
▪ Chemical treatments: alternatively, growth regulators or other chemicals may be applied to induced germination
growth regulators (use of plant hormones) commonly used GA3 (100ppm) and kinetin (10-15ppm).

C. Miscellaneous approaches
▪ Exposing seeds to white light: Photo dormancy or light sensitivity affects germination of some seeds.
▪ Pressure treatment: can be kept in autoclave and required pressure can be applied for breaking dormancy.
▪ Infra-red radiation treatment: Infra-red rays can be passed on to the seeds and dormancy can be released.
▪ Magnetic seed treatment: can be kept in the magnetic field for about 1 to 10 days for breaking dormancy
▪ Hot water treatment: It is effective in case of leguminous tree crop seeds. The seeds should be soaked in boiled
water for 1-5 minutes or 60-80 minutes.
▪ Micro-organism’s reaction: hard seed coat.

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 : SEED GERMINATION PHYSIOLOGY
4.1. Introduction
In seed germination process, the seed’s role is that of reproductive unit; it is the thread of life that assures the survival
of all plant species. Furthermore, because of its role in stand establishment, seed germination remains a key to modern
agriculture. Seed would normally germinate only after they have undergone a predefined period of growth and
development accumulating food reserves and finally becoming air dry. Seed germination depends on a favorable
combination of several external and internal factors; in nature, seed must wait for this combination to occur for their
germination.

Definition: Seed germination is the resumption of active growth of the embryo that results in the rupture of the seed
coat and the emergence of the young plant under favorable conditions. When strictly defined - Germination is the initial
emergence of the radicle from the seed coat.

4.2. Types of Germination

I. Hypogeal (hypogeous) germination. Type of germination in which the cotyledons remain below the ground while
the epicotyl elongates; only plumule emerges above the ground. e.g., Most of the monocots and pea
II. Epigeal (Epigeous) germination. Type of germination in which the cotyledons are forced above the ground by the
elongation of the hypocotyl. E.g., Most of the dicots (bean) and pine.
III. Vivipary
Germination of seed inside the fruit which is still attached to the mother plant and which also nourishes the seedling at
initial stages just after germination is termed as vivipary. The salinity of the surrounding soil damages the seeds if they
fall in their early stages. In such an environment, if seeds are dispersed then their chances of survival are very less.
Many plants growing along sea coasts e.g., mangroves.

IV. Pre-harvest Sprouting

Sprouting of seeds of standing crop, due to high moisture is known as pre-harvest sprouting. e.g., Groundnut.

V. Hypo-epigeal Germination
In the dicot species, one cotyledon shows hypogeal germination by remaining beneath the soil as in hypogeal
germination while the other cotyledon comes out above soil as epigeal germination. e.g., Peperomia peruviana

4.3. Phases of Seed Germination

Phase I. Imbibition: Rapid water uptake

▪ Germination begins with the absorption of water, which induces gibberellin synthesis by embryo.

Phase II. Active metabolism: Major metabolic events begin (continuation of respiratory activity).
▪ GA stimulates α-amylase production by aleurone (RNA & protein synthesis stimulated).
▪ Amylase breaks down starch reserves into maltose in endosperms (Hydrolysis (digestion) of food reserves by
enzymes)
▪ Maltose is either hydrolyzed (to glucose) for energy, or polymerized (to cellulose) for cell wall formation.

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 ▪ This energy & cellular building blocks are used to promote cell division & the growth of an embryonic shoot
(Induction of cell division & cell growth).

Phase III. Cell expansion: radicle extension and the completion of germination.
▪ Sugars fuel the growth of embryo.
▪ The seed coat (Testa) ruptures and the embryonic root (radicle) grows first into the ground to extract key
nutrients and minerals.
Post germination stage
▪ Controlled growth of root and shoot axis.
▪ Controlled transport of materials from food stores to growing axis.
▪ Senescence (aging) of food storage tissues.

Mobilization of stored reserves (seedling growth begin)

▪ Mobilization of the polymeric food reserves present within the storage is a post-germinative event
▪ The food reserves are converted in to readily transportable forms
▪ More food reserves are transported to rapidly metabolizing and growing organs of the seedling
▪ Reliance on the stored reserves diminishes as the seedling becomes photosynthetically active

The germination process requires that:

▪ The plant embryo leaves the quiescent state
▪ Mobilize stored nutrients
▪ Overcome the barrier of surrounding issues, and
▪ Resume cell elongation, cell division, and development

Figure: Structure of a barley grain and the functions of various tissues during germination

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▪ Respiration that supplies metabolic energy is activated immediately following imbibition’s
▪ Germination-related genes will be transcribed and translated into proteins within the 1st first few hours following
hydration
▪ Cell division only begins following the completion of germination
▪ Virtually all of the cellular and metabolic events that are known to occur before the completion of germination of
non-dormant seeds also occur in imbibed dormant seeds.
▪ Uptake of water by a mature dry seed is triphasic
- A rapid initial uptake (phase I)
- Followed by a plateau phase (phase II)
- A further increase in water uptake occurs only after germination is completed but dormant seeds do not complete
germination, they cannot enter phase III
▪ Germination sensu stricto (in the narrow sense) does not include seedling growth
▪ Seedling growth commences after germination is completed
▪ Extensive mobilization of the major storage reserves, are also not part of germination. Such events are post-
germination events.

Figure: Time course of water uptake and some important changes associated with germination and early
seedling growth. The time taken for these events to be completed varies among species and the germination
conditions to which the seed is subjected.

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4.4. Factors Affecting Seed Germination

Specific seed germination requirements vary depending on the plant species. There are internal and external factors:
I. Internal factor: Seed maturity, Dormancy, Mechanical damages
II. External factors: a. Water b. Oxygen c. Temperature d. Light e. Soil Factors

A. Water: Water is a basic requirement for germination. It is essential for enzyme activation, breakdown, translocation
and use of reserve storage materials. Seeds have 4-12 % of moisture in the packaged state. For germination, moisture is
increased to 25-50%.

B. Oxygen: atmospheric air is composed of 79.9 % N, 20% O2 and 0.03 % CO2. Oxygen is required for germination of
most of species. If CO2 concentration is higher than 0.03 % it retards germination. Respiration increases sharply during
seed germination. Since respiration is essentially an oxidative process, an adequate supply of oxygen is a must.

C. Temperature: Seed germination is a complex process involving many individual reactions and phases, each of which
is affected by temperature. The effect on germination can be expressed in terms of cordial temperature i.e., minimum,
optimum and maximum temperature. The optimum temperature for most of the seeds is between 15 to 30 0C maximum
temperature is between 30 to 400C. Some species will germinate even at freezing point. e.g., Alpine.

D. Light: Some species required light for seed germination. Both light intensity (lux) and light quality (colour and
wavelength) influence seed germination

E. Soil factor: Soil structure, soil texture and soil temperature influences on seed germination. seeds also require a
suitable soil pH in order to germinate (for optimal function of enzymes).

Roles of plant hormones with respect to seed germination and dormancy

For regulation of germination the promoters and inhibitors present in the seed should be in a balanced manner.
▪ Gibberellins: GA3 enhances germination. GA helps in translocation of food reserve materials to active site of
meristematic activity. GA also helps in cell division.
▪ ABA: Delay’s germination. Abscisic acid is an inhibitor that can prevent germination by affecting RNA synthesis.
▪ Ethylene (ETH): in particular, often exhibits a promotive effect on germination.
▪ Brassinosteroids (BRs): are another group of plant hormone that can promote germination
▪ Ethylene and brassinosteroids also influence the ABA signaling pathway
▪ Both Ethylene and brassinosteroids reduce the ability of ABA to inhibit germination
▪ Cytokinin: had a positive effect on seed germination Cytokinin is a natural endogenous hormone which controls
germination through DNA to RNA transcription system.

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 : SEED QUALITY TESTING
5.1. Seed Testing
The quality of seed cannot be assessed by visual observation, instead objective methods have been developed. Evaluation
of the seed quality is vital in order to predict its performance in the field and determine its value for planting. Seed
analysis laboratories are, therefore, now widespread. They adopt standardized methods to eliminate poor quality seed
lots and ensure improved results for farmers at harvest. The primary aim of the seed testing is:
➢ To obtain accurate and reproducible results regarding the quality status of the seed samples submitted to the seed
testing laboratories.

Historic Events of Seed Testing in the World

1816: - City of Bern, Switzerland, enacted legislation prohibiting the sale of adulterated clover seed, earliest regulation
on the sale of seed.
1869: - First seed testing laboratory established in Saxony, Germany, by Friedrich Nobbe.
1876: - E. H. Jenkins established the first seed testing laboratory in the United States.
1900: - In Europe there was 130 Seed Testing Stations
1924: - Formation and organization of the International Seed Testing Association.
1931: - The first set of international rules by the International Seed Testing Association (ISTA, 1931).
1939: - Passage of the Federal Seed Act in the United States (AOSA)
1972: - In Ethiopia the first Seed Testing Laboratory was established by CADU at Kulumsa.

Why seed testing is important?

Seed testing is an analysis of physical parameters and physiological qualities of a seed lot, based on a small representative
sample. “Quality” i.e., physiological quality, not genetic quality is a measure of potential performance under optimal
conditions. Seed testing is, therefore, an important tool for ensuring that farmers get the quality of seed they want.
Testing is also used in seed law enforcement to protect buyers from fraudulent sales and to provide technical professional
opinions in cases of litigation arising from differences between the information on certification labels and actual results.

For farmers, seed testing:

▪ Guarantees that seeds meet minimum quality standards in terms of physical purity and germination percentage.
▪ Minimizes the risk of crop failure; and avoids problems resulting from use of seed contaminated with noxious weeds,
seed infested with disease or insects, and seed having a low viability level.

Seed scientists and technologists have developed standard testing procedures to extract detailed information on those
quality characteristics that determine the value of seed. It is important that evaluation methods and test results are
consistent and uniform. For this reason, International Seed Testing Association (ISTA) guaranteeing worldwide annually
updated, harmonized and uniform seed testing methods.

Role of seed testing laboratory

The seed testing laboratory is central to the seed certification process; it must meet the standards set by seed legislation.
A seed laboratory does not improve the quality of seeds produced and distributed in the country; rather, based on the
results of tests on samples, it provides information to avoid or at least mitigate the adverse effects of using poor quality
seed.

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Procedures for laboratory seed testing
Sample reception and registration
When submitted samples arrive at the testing laboratory, but before registration, check the following:
▪ Samples are sealed.
▪ Two samples are provided (one for use as a working sample and the other for use in the post-control test).
▪ Samples are accompanied by sampling reports/forms.
▪ Sample bags are neither wet nor torn.
▪ Labels both inside and outside contain all the required information.

Register each sample received including:

▪ Label details
▪ Information in the sampling report
▪ Test identification number
▪ Sample reception date
▪ Test completion date
▪ Reference and nature of documents issued to the customer.

Computerized information systems are widely used in laboratories to facilitate data management. Assign a code to
identify the all samples taken from it for analysis. The simplest method is to give consecutive serial numbers, for
example: 0004 = the fourth sample received at the laboratory; or 19003 = the third sample received in 2019.

Prepare a test sheet including the following information:

▪ Registration number (code)
▪ Reception date
▪ Species, variety and seed class
▪ Requested test

Following registration, to proceed with testing, it is necessary to obtain a working sample by reducing the submitted
sample in the laboratory using appropriate methods.

5.2. Seed Quality Testing

5.2.1. Seed sampling
The purpose of seed sampling is to obtain a representative sample of a seed lot. This sample size must be such that
laboratory tests can determine the probability of occurrence of different constituents in the seed lot. Seed sampling
requires in-depth knowledge of the rules and methods. A properly trained seed sampler must take a good sample that
represents as accurately as possible the quality of the seed lot.

Seed lot sampling procedures

Seed testing is based on lots, i.e., precise quantities of seed. Seed lots should be uniform and harvested from a specific
seed field so that analysis results can be related to particular fields. The size of the seed lot depends on the size of the
seed. In general, the bigger the seed, the bigger the seed lot.

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I. Collecting a primary sample
A primary sample is a portion taken from the seed lot in the warehouse in a single sampling action. In order to meet
statistical requirements, ISTA defines the minimum number of primary samples for three different container types (In
the table below):
▪ < 15 kg
▪ 15-100 kg
▪ > 100 kg (or streams of seed entering containers)

Minimum sampling intensity for seed lots in containers

Weight of individual Weight of lot (kg or number of Number of primary samples
container in the seed lot containers)
≤ 500 kg ≥5
> 100 kg 501–3 000 kg 1 for each 300 kg (but ≥ 5)
3 001–20 000 kg 1 for each 500 kg (but ≥ 10)
20 001 kg > 1 for each 700 kg (but ≥ 40)
1–4 containers 3 from each container
15–100 kg 5–8 containers 2 from each container
9–15 containers 1 from each container
16–30 containers 15 from the seed lot
31–59 containers 20 from the seed lot
≥ 60 containers 30 from the seed lot
Containers shall be combined into smaller units ≤ 100 kg (20 containers of 5 kg, 33
Containers < 15 kg containers of 3 kg or 100 containers of 1 kg)
For sampling purposes, each sampling unit is regarded as "one container"
Sampling intensity is as defined for containers for 15-100 kg

II. Obtaining a composite sample

The composite sample is formed by combining and mixing all the primary samples taken from the seed lot. To obtain a
composite sample, it is necessary to combine and mix all the primary samples taken from the seed lot. During sampling,
primary samples are compared to check for homogeneity. If the primary samples appear uniform, they are combined to
form the composite sample. Otherwise, the sampling procedure is interrupted.

III. Obtaining a submitted sample

A submitted sample is a sample sent to the testing laboratory. It may comprise the entire composite sample or a
subsample obtained using one or more ISTA reduction methods as for the working sample.

The packaging depends on the specific test requirements. Suitable containers include an unused clean cloth, a paper
bag or good quality manila envelope. When the sample is to be tested for moisture content, a moisture-proof container
must be used. Samples should never be left unprotected or exposed to moisture, heat or direct sunlight. Containers are
sealed to discourage tampering and labels are attached both inside and outside the bag.

There is a fixed minimum weight for submitted samples depending on the species and according to the tests requested.
It is important to forward the submitted sample to the seed-testing laboratory in a safe and timely manner, together with
the sampler report. The sampler must always keep a copy of the sampling report.

IV. Obtaining a working sample

A working sample is obtained in the laboratory from the submitted sample using an appropriate reduction method. It is
the working sample that actually undergoes seed testing.

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Methods for obtaining working sample
The working sample must be representative of the submitted sample. Mix thoroughly the submitted sample before
dividing it mechanically or manually.
▪ Mechanical methods (not appropriate for seed health testing): Soil divider, Centrifugal divider Conical
divider, Rotary divider and Variable divider.
▪ Manual methods: Spoon method, Modified halving and Hand halving method

Seed sampling instruments

A. Sampling by hand
This method is appropriate for all species. Moreover, it is potentially the most suitable method for seed that could get
damaged using triers, seeds with wings, seeds with low moisture content, seed tapes and seed mats.

B. Nobbe trier
The Nobbe trier or dynamic spear is a pointed tube with an opening near the pointed end. Seed passes through the tube
and is collected in a container.

C. Sleeve-type trier
The sleeve-type trier – stick trier or sampling stick – consists of an inner and an outer tube. Some models have partitions,
others have only one cavity. The ISTA Rules specify that a stick without partitions can only be used horizontally.

D. Automatic sampling from a seed stream

Automatic sampling devices are available. It is important to ensure that: a uniform sample is taken from the whole cross-
section of the seed stream.

Operation can be either manual or automatic. The intervals between taking primary samples should be constant;
however, samples may also be taken at random intervals.

5.2.2. Physical purity analysis

Objectives
▪ Determine percentage composition by weight of the sample being tested (and by inference the composition of the
seed lot).
▪ Identify the various species of seeds and inert particles constituting the sample.
It is essential to follow all the general requirements established in the regulations. The working sample is divided into
three components: Pure seed; Other seeds and Inert matter.

Identify all species of seed and each kind of inert matter present, and then determine the percentage of each part by
weight.

I. Pure seed: the species stated by the applicant, or that found to predominate in the test, and shall including all
botanical varieties and cultivars of that species. The pure seed fraction comprises also:

- By definition, the pure seed shall include, immature, undersized, shriveled, diseased or germinated, providing they
can be identified as of the species, unless transformed into visible fungal sclerotia, smut galls or nematode galls.; and
Intact seeds as defined by each genus; e.g., florets with an obvious caryopsis containing, free caryopses as in
Gramineae or pieces of seed units larger than one half of the original size.

II. Other seeds: these include seed units of any plant species other than of pure seed.

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III. Inert matter: this includes seed units and all other matter and structures not defined by ISTA as pure seed or
other seed, for example:
- Pieces of broken or damaged of pure seed unit’s half or less than half the original size.
- Seeds of Fabaceae(legumes) with the seed coat entirely removed and separated are considered as inert matter.
- Soil particles, sand, stones, chaff, stems, leaves, flowers and smut balls, ergots, nematode galls and all other non-seed
matter.
Apparatus
▪ Purity board: the main instrument.
▪ Hand lenses and binocular microscopes: often used for accurate identification and separation of small seed units
and fragments.
▪ Seed blowers: mechanical devices used to separate light weight material (e.g., chaff and empty florets) from heavier
seeds.
▪ Sieves: used to separate components of different sizes into sized fractions. Examine each fraction and classify the
particles (pure seed, other seed or inert matter).
▪ Analytical balance, forceps and fine needles.

Procedure:
▪ Purity is conducted using a working sample obtained from a submitted sample.
▪ Determine the correct weight of the working sample (≥ 2 500 seeds with a maximum weight of 1 000 g). The ISTA
Rules stipulate that the analysis can be done on one working sample of this weight or on two subsamples of at least
half this weight, each independently drawn.
▪ Weigh the working sample (or each subsample) in grams.

Divide the working sample on the working board into three components (pure seed, other seed and inert matter).
Weigh the individual fractions independently using an analytical balance.
For example: - Pure seed = X (g)
- Other seeds = Y (g)
- Inert matter = Z (g)

Calculation and expression of results

• Total weights of all the components and subtract from weight of working sample.
• If there is a discrepancy of >5% retest sample.

Add together percentages of all fractions. Fractions that are to be reported as trace should be excluded from the
calculation added.

All the three components must be weighed to the required number of decimal places. The percentages of the
components are determined as follows

% Of component = Weight of individual component (X) x 100

Total weight of all components (X+Y+Z)
If there is a gain or loss between the weight of the original samples and the sum of all the components is in excess of
one percent, another analysis should be made.
If all replicates of all fractions are within tolerance, add the weights of the corresponding fractions together, calculate
percentages and round the figures off to one decimal place.

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5.2.3. Germination test
The objective of the germination test is to determine the germination potential of a seed lot, which can then in turn be
used to compare the quality of different lots and also estimate the field planting value.

Testing under field conditions is normally unsatisfactory, as the results cannot be repeated with reliability. Laboratory
methods have, therefore, been evolved in which the external conditions are controlled to give the most regular, rapid
and complete germination for the majority of samples of a particular species. The conditions have been standardized to
enable the test results to be reproduced within limits as near as possible to those determined by random sample variation.

Definitions: Germination of a seed in an ISTA test is the emergence and development of the seedling to a stage where
the aspect of its essential structures indicates whether or not it is able to develop further into a satisfactory plant under
favorable conditions in the field.

Germination is defined as the emergence and development from the seed embryo, of those essential structures, for the
kind of seed in question, indicates its ability to produce a normal plant under favorable conditions.

The germination percentage indicates the proportion by number of seeds which have produced seedlings classified as
normal under favorable conditions and within the specified period i.e., the percentage of normal seedlings.

I. General principles
Germination tests must be made with pure seeds. The pure seed definition for the species must be applied. The pure seed
can be taken from either the pure seed fraction of a purity test or from a representative fraction of the submitted sample.
A minimum of 400 seeds are required in four replicates of 100 seeds each or 8 replicates of 50 seeds each or 16 replicates
of 25 seeds each depending on the size of seed and size of containers of substrate.

The test is conducted under favorable conditions of moisture, temperature, suitable substratum and light if necessary.
No pretreatment to the seed is given except for those recommended by ISTA.

The seeds are spread apart on the substratum to prevent seedlings from coming into contact with one another before they
are counted and removed. At the end of the specified germination period, the replicates are examined; two counts are
usually made of the seedlings and seeds in each category undergoing testing.

Methods for germination tests for some species (ISTA, 2016)

Species Substrate temperature (°C) First Final Recommendations
count count (d) for breaking
(d) dormancy
Barley - Hordeum vulgare L. BP; S 20 4 7 Preheat at 30-35°C;
GA3; KNO3; prechill
Groundnut -Arachis BP; S 20 <=> 30; 25 5 10 Remove shells,
hypogaea L. Preheat at 40±2°C;
Maize - Zea mays L. BP; TPS; S 20 <=> 30; 30; 25 4 7
Sorghum – Sorghum bicolor TP; BP 20 <=> 30; 25 4 10 Prechill
(L.) Moench
Wheat – Triticum aestivum TP; BP; S 20 4 8 Preheat at 30-35°C;
L. GA3; prechill
st nd
* The symbol "<=>" indicate alternating temperature regimes. 1 temperature: 16 h; 2 temperature: 8 h BP= between paper, S =
Sand, TP = Top of paper, TPS = top of paper or sand. Preheat at 30-35°C; prechill

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The germination percentage reported on the certificate issued indicates the proportion by number of seeds that have
produced “normal” seedlings under the conditions and within the period specified by the Rules.

Essential seedling structures

A seedling, depending on the species tested, consists of a specific combination of some of the following structures
essential for its development into a satisfactory plant:
▪ Root system (primary root; in certain cases, seminal roots)
▪ Shoot axis (hypocotyl; epicotyl; in certain Poaceae mesocotyl; terminal bud)
▪ Cotyledons (one or more) • Coleoptile (in all Poaceae)

The 50 % rule
▪ The 50% rule is used to evaluate cotyledons and primary leaves.

Cotyledon tissue:
▪ Seedlings are “normal” when at least half the total cotyledon tissue is functional.
▪ Seedlings are “abnormal” when more than half the cotyledon tissue is missing, necrotic, decayed, discolored.

Primary leaves (evaluated in species such as Phaseolus):

▪ Seedlings are “normal” when at least half the primary leaf tissue is functional.
▪ Seedlings are “abnormal” when more than half the primary leaf tissue is missing, necrotic, decayed or discolored.
II. Growing media
Growing media must provide sufficient pore space for air and water, growth of the root system and contact with solutions
(water) necessary for plant growth. The prescribed media are sand and paper and each has advantages and disadvantages.
Sand provides a more natural environment for seedling growth. The contact between the seed and sand is good, the
shoots grow upwards and emerge from the media, and the seedling is exposed to light which allows improved
development of essential structures. However, it requires more space in germination chambers than paper substrata, it is
heavy to move during testing and at disposal, and it requires significant storage space. The laboratory can be designed
to minimize these concerns. Sand is often the preferred media when retesting due to fungal infection, or because of any
condition rendering seedling evaluation difficult in a paper substrate. In these cases, it may be necessary to sterilize the
sand. Sand used as growing media must have ≥ 90% of the particles should pass through a sieve with holes or meshes
of 0.8 mm width, and be retained on a sieve with holes or meshes of 0.05 mm width.

A. Paper growing media

Methods using paper:
▪ Top of paper (TP) – seeds germinate on top of one or more layers of paper.
▪ Between paper (BP) – seeds germinate between two layers of paper.
▪ Pleated paper (PP) – seeds are placed in a pleated, accordion-like paper strip with 50 pleats, usually two seeds to
each pleat. This method is an alternative to TP and BP.

The paper must be prepared from wood, cotton or other purified vegetable cellulose. The paper may take the form of
filter papers, blotters or towels. The paper should be such that:
▪ The roots of the seedlings will grow on and not into it;
▪ It possesses sufficient strength to enable it to resist tearing when handled during the test.

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B. Sand or organic growing media
Methods using sand or organic growing media:
▪ Top of sand (TS), top of organic growing medium (TO) – seeds are pressed into the surface of the sand or the
organic growing medium.
▪ Sand (S), organic growing medium (O) – seeds are planted on a level layer of moist sand or organic growing
medium and covered with 10–20 mm of uncompressed substrate, depending on the size of the seed.

The following elements should be used in known proportions and fitting the requirements for sand or organic media:
▪ Mineral particles: for example, sand, perlite and vermiculite. The proportion should be around 20 % in volume. It
is recommended that 90 % of the particles should pass through a sieve with holes of 2 mm width and be retained on
a sieve with holes or meshes of 0.05 mm width.
▪ Organic compounds: fibers such as peat, coconut fibers or wood fibers, with a size less than 5 mm.

C. Soil
Soil is generally not recommended as a primary growing medium. However, it may be used as an alternative to organic
growing media when seedlings show phytotoxic symptoms or if evaluation of seedlings is in doubt on paper or sand.

III. Apparatus
▪ Bell jar or Jacobsen apparatus (Copenhagen tank)
▪ Germination incubator and room germinator
▪ Petri dishes, forceps, covering net, blotting paper, and sand.

IV. Procedure (TP):

▪ Take a sample of 400 seeds at random from well-mixed pure seed. It is important to not select seeds, as this would
give biased results.
▪ Use four replicates of 100 seeds to ensure adequate spacing. Split replicates of 50 seeds (or even 25, particularly
where there are seed-borne pathogens or saprophytes present) to minimize the effect of adjacent seeds on seedling
development.
▪ Place seeds uniformly and sufficiently apart on the moist substrate on the Petri dish. If seeds grown on paper
substrates are heavily infected, at an intermediate count, transfer remaining seeds and seedlings to fresh media.
▪ Place Petri dishes in the germination apparatus; record the number of seeds set and the date.
▪ Make two counts of seedlings. Schedule the first and final counting’s according to the ISTA Rules.
▪ Keep the seed moist throughout the test period.

The germination percentage is expressed as follows:

Germination (%) = Number of normal seedlings × 100

Number of seeds set for germination

Seedlings are evaluated and categorized as follows:

A. Normal seedlings
Normal seedlings show the potential for continued development into satisfactory plants when grown in good quality soil
and under favorable conditions of moisture, temperature and light.

To be classified as normal a seedling must conform to one of the following categories:

▪ Intact seedlings: seedlings with all their essential structures well developed, complete, in proportion and healthy.

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▪ Seedlings with slight defects: seedlings showing certain slight defects of their essential structures, they show an
otherwise satisfactory and balanced development comparable to that of intact seedlings of the same test.
▪ Seedlings with secondary infection: seedlings which it is evident would have conformed to one of the above, but
which have been affected by fungi or bacteria from sources other than the parent seed.

B. Abnormal seedlings
Abnormal seedlings do not show the potential to develop into a normal plant when grown in good quality soil and under
favorable conditions of moisture, temperature and light. The following seedlings are classified as abnormal:

▪ Damaged seedlings: seedlings with any of the essential structures missing or so badly and irreparably damaged that
balanced development cannot be expected;
▪ Deformed or unbalanced seedlings: seedlings with weak development or physiological disturbances or in which
essential structures are deformed or out of proportion;
▪ Decayed seedlings: seedlings with any of their essential structures so diseased or decayed as a result of primary
infection that normal development is prevented.
C. Ungerminated seeds
Seeds which have not germinated by the end of the test period, when tested under favorable conditions are classified as
follows:

Hard seeds: seeds which remain hard at the end of the test period, because they have not absorbed water. Hard
seededness is a form of dormancy. It is common in many species of the Fabaceae but may also occur in other families.

Fresh seeds: seeds, other than hard seeds, which because of dormancy have failed to germinate under the conditions of
the germination test, but which remain clean and firm and have the potential to develop into a normal seedling. When 5
% or more of fresh seeds are believed to be present, their potential to germinate must be determined by dissection,
tetrazolium or excised embryo.

Dead seeds: seeds which at the end of the test period are neither hard nor fresh nor have produced any part of a seedling.
If it can be seen that a seed has produced any part of a seedling (e.g., the tip of the primary root) even though decayed
at the time of assessment, it is counted as an abnormal seedling and not as a dead seed.

Other categories: Upon request of the customer, the number of empty, embryo less or insect-damaged seeds may be
determined and reported under ‘Other Determinations’ on the ISTA Certificate.

V. Retesting
The result of a test must be considered unsatisfactory and must not be reported, and a second test must be made by the
same or an alternative method, under the following circumstances:
▪ When dormancy is suspected (fresh ungerminated seeds).
▪ When the result may not be reliable because of phytotoxicity or spread of fungi or bacteria.
▪ When there is difficulty in deciding the correct evaluation of a number of seedlings.
▪ When there is evidence of errors in test conditions, seedling evaluation or counting.
▪ If a sample does not respond satisfactorily to the methods elected.
▪ When the range for the replicates exceeds the maximum tolerated range, a retest must be carried out using the
same test method.

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VI. Calculation and expression of results
The result of the germination test is expressed as a percentage by number of normal and abnormal seedlings and hard,
fresh and dead seeds. The percentages are rounded to the nearest whole number. For example, 75.00 and 75.25 are
rounded to 75%; 75.50 and 75.75 are rounded to 76%. The sum of the percentages of normal and abnormal seedlings
and ungerminated seeds must be 100.

VII. Tolerances
The result of a germination test can be relied upon only if the difference between the highest and the lowest replicates
is within accepted tolerances. To check the reliability of a test result, the average percentage of the replicates is rounded
to the nearest whole number and compared with tolerance table. The result is considered reliable, if the difference
between the highest and the lowest replicate does not exceed the tolerance range. Tolerances are applied to at least the
category of normal seedlings. If the range of the replicates exceeds the maximum tolerated range a retest must be made.

5.2.4. Viability test using tetrazolium

Tetrazolium test is a biochemical test and one of the quick methods to predict seed viability. Viability is the capability
of the seed to germinate and produce a normal seedling. It indicates that a seed contains the structures and substances
required to germinate under favourable conditions in the absence of dormancy. External physical appearance alone
cannot determine whether a seed is alive or dead. Seed viability testing is therefore carried out to determine the
percentage of viable seed in a given lot. The test is valid for all species for which a method is described in the ISTA
Rules.

Objective: Make a rapid assessment of seed viability and seed vigour in the following circumstances:
▪ Seeds need to be sown shortly after harvest.
▪ Seeds have deep dormancy or show slow germination.
▪ A very quick estimate of germination potential is required (when a seed lot is received at a processing plant).
▪ A solution is required to problems encountered in a germination test (e.g., reasons for abnormals are not clear or
treatment with pesticides is suspected).

I. General principles
The tetrazolium test distinguishes between viable and dead tissues of the embryo on the basis of their relative respiration
rate in the hydrated state. Although many enzymes are active during respiration, the test utilizes the activity of
dehydrogenase enzymes as an index of the respiration rate and in turn the seed viability.

The tetrazolium test distinguishes between viable and dead tissues of the embryo on the basis of their relative respiration
rate in the hydrated state. Although many enzymes are active during respiration, the test utilizes the activity of
dehydrogenase enzymes as an index of the respiration rate and in turn the seed viability.

In the topographical tetrazolium test a colorless solution of 2,3,5-triphenyl tetrazolium chloride or bromide is used as an
indicator to reveal the reduction processes which take place within living cells. The indicator is imbibed by the seed.
Within the seed tissues it interacts with the reduction processes of living cells and accepts hydrogen from the
dehydrogenases. By hydrogenation of the 2, 3, 5-triphenyltetrazolium chloride a red, stable and non-diffusible substance,
triphenyl formazan, is produced in living cells. This makes it possible to distinguish the red-colored living parts of seeds
from the colorless dead ones.

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A viable seed shows staining in all those tissues whose viability is necessary for normal seedling development. Seeds
can then be classified into viable and non-viable seed classes.

II. Apparatus: Petri dishes, filter paper, magnifying glass, dropper and bottle, solution of tetrazolium, dissecting
needles, forceps.

III. Procedure
▪ Draw four replicates of 100 pure seeds at random, either from the pure seed fraction or submitted sample.
▪ Mix the pure seed fraction thoroughly taking care to not select seeds causing biased results.
▪ Soak seeds in water overnight to soften the embryo and endosperm and activate the enzyme system.
▪ Make a cut or completely remove the seed-coat. e.g., dicots, bisect longitudinally: e.g., Sorghum, large seeded grasses,
bisect laterally. e.g., Small seeded grasses and conditioning only. e.g., large seeded legumes – to expose the embryo
and facilitate contact with the tetrazolium solution. This process known as seed preparation
▪ Immerse the prepared seeds or embryos in tetrazolium salt solution. Avoid exposure to direct light, as it would cause
a reduction of the tetrazolium salt. Refer to the ISTA Rules for optimum temperatures and staining times.
▪ Wash seeds repeatedly with distilled water.
▪ Examine seeds under a magnifying glass.

IV. Staining
There are optimum temperatures and staining times. Staining temperatures used may deviate but must be in the range
of 20–40 °C. If the optimum staining temperature of 30 °C is not used, then suitable adjustments in staining duration
must be made, as an increase/decrease of 5 °C from the optimum of 30 °C reduces/increases staining time by one half.
Staining periods should not be taken as absolute, because they may vary according to the condition of the seed.

Example: For maize, the seeds are soaked for 18 hours in water at 20 °C, then cut longitudinally through the embryo
and ¾ of the endosperm before staining in 1% tetrazolium solution for 2 hours at 30 °C. The seeds are then halved and
the cut surfaces observed.

Seeds are classified as follows:

▪ Living – red or purple embryo.
▪ Dead – no colour in the embryo. Even when the embryo is not stained but red colour develops on other parts, the
seed is still classified as dead.

V. Calculation, expression of results and tolerances

Maximum tolerated ranges for replicate differences are the same as for germination tests.

The percentage of viable seed is calculated as follows:

Viable seed (%) = Number of living seeds x 100

Number of seeds set for viability test

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5.2.5. Determination of seed moisture content
Moisture content is crucial for preserving quality of stored seed and maintaining viability. Seeds with the correct
moisture content can be stored longer and are relatively resistant to damage by insects.

The moisture content of a sample is derived from the loss in weight when the seed is dried in accordance with ISTA
methods. It is expressed as a percentage of the weight of the original sample. The ISTA Rules differentiate between
direct and indirect methods to determine moisture content. Direct methods involve oven drying, desiccation and other
physicochemical approaches. Note that some species require grinding before testing.

Seed it has hygroscopic nature, so it gains or loses moisture depending on the humidity of the air around it. So, a
moisture test taken today may bear little relevance to the moisture of the same seed lot some days or weeks later.

Objectives:
▪ Determine the appropriate drying conditions for optimum preservation during storage.
▪ Verify whether seed complies with the maximum moisture content specified in the seed regulations.

Methods of seed moisture determination

I. Air oven method
In this method, seed moisture is removed by drying the seed sample at a specified temperature for a specified duration
known as constant temperature oven method.

Constant temperature oven method is a direct method, it reduces oxidation, decomposition and loss of other
volatile substances, but ensures the removal of as much moisture as possible. The temperature (high /low constant)
depends on the species.

A. Low constant temperature method

The working sample must be evenly distributed over the surface of the container. Weigh the container and its cover
before and after filling. Place the containers rapidly on top of the cover in an oven maintained at a temperature 103±
2oC and dry for 17± 1hours. The drying period begins at the time the oven returns to the required temperature. At the
end of the prescribed period cover the container and placed it in a desiccator to cool for 30 – 45 minutes. After cooling,
weigh the container with it cover and content. Seeds containing high percentage of oil should be dried at 103°C for 17
hours. This method is suitable for oily seeds such as onion, capsicum, soybean, radish, groundnut, castor, mustard,
sesame, cotton etc.

B. High constant temperature method

The same as above except when temperature ranges between 130 to 133oC, most species are dried for 1 hr. at 130° C,
cereals for 2 hours (130°C) and maize for 4 hours (130°C).

Weighing working sample: Weighing shall be in grams to three decimal places.

Working sample: The determination shall be carried out in duplicate on two independently drawn working samples,
each of the following weight, depending on the diameter of the containers used: If less than 8cm diameter 4 – 5g and
8cm diameter or larger 10g. Before the working sample is drawn, the submitted sample shall be thoroughly mixed.

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Cutting: Large tree seeds (less than 5000 seeds/kg) and tree seeds with very hard seeds coats such as leguminous
species may be cut into small pieces instead of ground. The cutting shall be done as a sub-sample before drawing
working sample.

Grinding: Large seed must be ground before drying unless their high oil content makes them difficult to grind or
(particularly in seed such as linen with oil of high iodine number) liable to gain in weight through oxidation.

The grinding shall be done on a sub-sample before drawing the working.

Pre-drying: If the species is one for which grinding is necessary and the moisture content is more than 17% (or 10%
in case of glycine max and 13% in the case of Oryza sativa).

Apparatus required for air oven method: Grinding mill, electrically heated oven, containers, thermometer,
desiccator, suitable desiccant (e.g., silica gel), analytical balance, sieves, cutting tool.

Procedure
▪ Distribute the working sample evenly over the surface of the container (The submitted sample shall be accepted for
moisture determination only if it is in an intact, moisture proof container from which as much as air as possible has
been excluded).
▪ Weigh the container and its cover before and after filling.
▪ Quickly place the container on top of or next to its cover, in an oven.
▪ At the end of the prescribed period, cover the container, place in a desiccator (containing desiccant) and leave to
cool at ambient temperature.
▪ Once cooled, weigh the container with its cover and contents.

The moisture content as a percentage by weight is calculated to three decimal places for each replicate using the
following formula:

MC (%) = Loss of weight x 100 = M2 – M3 x 100

Initial weight M2 – M1
Where:
• M1 = weight (g) of container and cover
• M2 = weight (g) of container, cover and contents before drying
• M3 = weight (g) of container, cover and contents after drying

II. Moisture meter /Digital moisture meter method

Moisture meters estimate seed moisture quickly but the estimation is not as precise as by the air oven method. Moisture
estimation is made quick by the advent of digital moisture meters. The principle involved is that electrical conductivity
of moist material is directly proportionate to the amount of moisture content in it.

Moisture meter method is an indirect method, any type of moisture meter can be used, as long it meets the calibration
and determination requirements. The moisture content measured is calibrated against the moisture content obtained
using the oven method. Meters are very practical and particularly useful when a rapid result is required, for example,
when seed arrives at the processing plant after harvest and it is necessary to decide whether further drying is required.

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5.2.6. Cultivar or Varietal purity test / Genetic purity
Genetic purity means plants and seeds confirming to the characteristics of the variety as described by the breeder. Over
the years, this purity is lessened in a variety due to reasons like mutations, mechanical mixture and natural crossing
etc. One basic aim is to produce seed which is genetically pure to the set standards. Hence, it is essential to control the
factors which will deteriorate genetic purity.

The objective of species and variety verification is to determine the extent that the submitted sample conforms to the
species or variety as requested by the applicant, using methods not permissible in a purity test.

Varietal or species purity is tested in the laboratory, in the greenhouse, and in field plots. However, variety identification
in the laboratory is not standard practice, because most species have insufficient morphological features for accurate
identification. The best way to ensure varietal purity is field inspection (as part of the certification process “post-
control”).

Traditional varietal purity tests in the laboratory (e.g., visual examination of seeds and seedlings and chemical tests) are
the first step towards identifying a variety or narrowing down the range of possibilities. The morphological and
physiological characteristics of seeds, seedlings and plants are examined. In all tests, authentic samples of the
species/variety should be available for comparison.

An authentic standard sample is a seed sample of a known species or variety or a sample with a known specific trait.
It is recommended that this sample is of a known origin, e.g., a certified reference sample or a sample taken by an official
or another person who can ensure the sample identity and characteristics. This sample will be used for obtaining
enzymatic, protein or DNA profiles.

A standard reference is a valid descriptive attribute of a species or variety, e.g., zygosity, or an isozyme, protein or
DNA banding pattern produced by gel electrophoresis or similar techniques, or an allelic profile or nucleotide sequence
or a molecular weight standard (MWS) for protein or DNA. This trait should be obtained by a validated method and
should be from an authentic standard sample or obtained from a reliable source as for MWS.

General principles
Field of application: The determination of a species or variety is valid only if the species or variety is stated by the
applicant and an authentic standard sample of the species or variety is available for comparison to ensure the certainty
of the determination. The traits compared may be morphological, physiological, cytological or chemical.

Testing principles: The determination is carried out, depending on the species or variety in question on seeds, seedlings
or more mature plants grown in a laboratory, a glasshouse, a growth chamber or a field plot. When an authentic standard
sample is available, the working sample is compared with the authentic standard sample. Whenever possible, the
working sample and the authentic standard sample must be handled in the same way, e.g., in field plots they must be
grown contemporaneously, near each other and in identical environmental conditions, and the evaluation must be done
at the same stage of development. When a standard reference is used in a test, the interpretation of the result is done by
comparing the traits of the seeds, seedlings or plants of the working sample with the standard reference.

In the case of species or variety that are sufficiently uniform in one or more traits (e.g., in self-pollinated species), the
conformity of the working sample with an authentic standard sample can be determined and if possible, the degree of
conformity may be quantified. If the species or variety is not sufficiently uniform (e.g., in cross-pollinated species), the
proportion of any obvious off-types is calculated and the conformity of the working sample is expressed.

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Types of Species and variety tests
1. Examination of seeds
There may be different procedures for examining seeds.
For testing morphological traits, the seeds must be examined with the aid of a suitable magnifying apparatus when
necessary. For testing colour traits, the seeds may be examined under full daylight or light of limited spectrum, e.g.,
ultra-violet.
For testing chemical traits, the seeds must be treated with the appropriate reagent and the reaction of each seed must
be noted.
For testing bio-molecular traits, DNA, RNA, protein or other specific metabolic products are extracted from the seeds
and the traits may be detected, elucidated and quantified.

2. Examination of seedlings
The seeds must be germinated on an appropriate medium. When the seedlings have reached a suitable stage of
development, they are examined in whole or in part, with or without further treatment. For testing bio-molecular traits,
DNA, RNA, protein or other specific metabolic products are extracted from the seedlings and the traits may be detected,
elucidated and quantified.

3. Examination of plants in glasshouse or growth chamber

The seeds must be sown in suitable containers and maintained in environmental conditions necessary for the
development of the traits.

4. Examination of plants in field plots

When plants are tested in field plots, each working sample must be sown in at least two replicate plots. Observations
must be made during the whole growing period, but particularly during the time after earing (cereals) or the beginning
of flowering (other species) that the most distinct differences between plants of individual varieties become apparent;
consequently, it is during these periods that each individual plant should be examined.

5.2.7. Seed vigour test

Seed vigour tests are laboratory methods employed to evaluate the seed vigour, which is a complex phenomenon
involving biosynthesis of energy and metabolic compounds which is normally expressed in speed and totality of
germination, mechanical rupture, force of seedlings, tolerance of seed to environmental stresses and disease resistance.

The objective of a seed vigour test is to provide information about the planting value in a wide range of environments
and/or the storage potential of seed lots. The test provides additional information to the standard germination test to
assist in the differentiation of seed lots of acceptable germination.

A seed vigour test is either a direct or an indirect analytical procedure to evaluate the vigour of a seed lot under
standardized conditions.
A. Direct tests reproduce environmental stresses or other conditions in the laboratory, and the percentage and/or rate of
seedling emergence are recorded.
B. Indirect tests measure other characteristics of the seed that have proved to be associated with some aspect of seedling
performance.

Seed vigour testing is based on the fact that seed deterioration (by which vigour is lost) and seed viability losses occur
at different rates. Seed viability and vigour are both high and similar. However, while seed viability has decreased only
slightly, vigour has decreased significantly. The causes of poor seed vigour are many and include nutrition provided by,

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and position on the mother plant, environmental and climatic conditions during crop growth, stage of maturity at harvest,
postharvest treatments and genetic effects i.e., practically all aspects of crop production.

Acceptable germination: A seed lot of acceptable germination is one which, in the absence of seed dormancy, has an
acceptable standard germination level for that species.

The germination test does not detect quality differences among seed lots with similar germination percentages. A
vigour test is more sensitive and able to detect such differences.

It is possible to identify lots more likely to perform well under non-optimal environmental conditions. Indeed,
germination tests take place under optimal germination conditions and the results express the germination potential. This
figure may be quite different from actual performance under stressed field conditions. For example, two seed lots may
have similar germination potential (> 90%), but significant differences in seed vigour (In the table below). An efficient
vigour test must pinpoint differences in physiological potential not detected by viability tests and rank lots according to
performance potential.
Example of germination and emergence in two seed lots
Seed lot Germination Seedling emergence (%)
(%)
Field 1 (near ideal conditions) Field 2 (unfavorable conditions)
A (high vigour) 90 88 75
B (low vigour) 90 87 50
Field 1 – the stands of both high (A) and low (B) vigour seed lots are similar to their germination.
Field 2 – the low vigour seed lot (B) has poor seedling emergence compared with the high vigour seed lot (A). In
addition, despite seed lot A’s superior vigour, emergence in Field 2 is lower than the germination percentage;
therefore, stand may be unacceptable if conditions are stressful.

I. General principles
A vigour test assesses, either directly or indirectly, the physiological and physical basis of potential seed lot performance
in a wide range of environments, and provides a more sensitive differentiation between seed lots of acceptable
germination than does the germination test. Such information can be used to make informed decisions regarding the
value of different seed lots.

Vigour tests are able to provide:

▪ a more sensitive index of seed quality than the standard germination test.
▪ a consistent ranking of seed lots of acceptable germination in terms of their potential physiological and physical
quality.
▪ information on emergence and storage potential of seed lots to plan marketing strategy.

ISTA validated four types of vigour test methods

Vigour test Scope and field of application
Conductivity Pisum sativum (garden pea only, excluding petit-pois varieties)
Phaseolus vulgaris, Glycine max
Accelerated ageing Glycine max
Controlled deterioration Brassica spp.
Radicle emergence test Zea mays

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Conductivity test: Measurement of the electrical conductivity of leachates provides an assessment of the extent of
electrolyte leakage from plant tissues. Conductivity measurement of the soak water in which a bulk sample of seeds has
been steeped gives an estimate of seed vigour. Seed lots with high electrolyte leakage, i.e., high leachate conductivity,
are considered to have low vigour due to low membrane integrity, whilst those with low leakage (low conductivity) are
considered to have high vigour.

The accelerated ageing (AA) stress test exposes seeds for short periods to high temperature and high relative humidity
(≈ 95 %). During the test, the seeds absorb moisture from the humid environment and the raised seed moisture content,
along with the high temperature, causes rapid seed ageing. High vigour seed lots will withstand these extreme stress
conditions and age more slowly than low vigour seed lots. Thus, after AA, high vigour lots retain a high germination,
whilst that of low vigour lots is reduced.

The controlled deterioration (CD) test exposes seed to a high temperature while at a specified and constant raised seed
moisture content. These conditions cause seeds to deteriorate, or age, rapidly. The moisture content of a seed sample is
raised before the seeds are placed at the raised temperature, thus ensuring that all samples tested are exposed to a
predetermined degree of deterioration during the test. High vigour seeds retain a high germination after deterioration,
while the germination of low vigour seeds is reduced.

Radicle emergence (RE) test: A slower rate of germination is an early physiological expression of seed ageing, the
major cause of reduced vigour. The rate of germination of Zea mays is accurately reflected in a single count of radicle
emergence early in germination and this single count relates closely to other expressions of the rate of germination. High
counts of radicle emergence early in germination are indicative of high seed vigour; low counts indicate low seed vigour.

II. Procedures
A. Working sample: The required number of seeds and replicates must be taken at random from the pure seed.

B. General directions: Different vigour test methods (direct and indirect) are described under three general categories:
▪ Stress tests – cold, accelerated ageing and controlled deterioration tests
▪ Biochemical tests – conductivity and tetrazolium tests
▪ Seedling growth and evaluation tests

The vigour index was computed by formula suggested by Abdul Baki and Anderson (1973).
Vigour index-I = (Average root length + Average shoot length) × Germination %
Vigour index-II = Average dry seedlings wt. (g) × Germination %

C. Test conditions: Methodology for each test is prescriptive, and no other methodology may be used if an ISTA
Certificate is issued.
D. Control samples: All vigour tests require rigid control of test conditions and, where specified, should include a
control seed sample to provide internal quality control of vigour test uniformity. Variability in control seed sample
results provides an indication of slight fluctuations in test conditions (e.g., changes in temperature or seed moisture)
which can significantly affect the reliability of results.

5.2.8. Seed health test

Seed health is the presence or absence of disease-causing organisms (e.g., fungi, bacteria and viruses) and animal pests
(e.g., nematodes and insects). Insects, pathogens and nematodes may affect the seed so as to make it ungerminable. Seed
borne inoculums may also give rise to progressive disease development in the field and reduce the yield of the crop.

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Seed health concerns the overall condition of seeds in terms of pathogenic infection of seeds, insect infestation,
morphological and physiological disorder (s) etc.

The objective is to determine the health status of the seed sample i.e., whether the seeds are infected with fungi or not.
Seed borne inoculum may give rise to spread of disease in field and reduce the commercial value of the crop. Further
the seeds imported from outside source may introduce new pests and diseases into new areas.

Methods for seed health testing

The methods adopted for seed health testing depends upon the type of disease/ pest infestation being tested. Some of the
tests adopted are given below

1. Visual examination: the seed sample is examined with the naked eye or hand lens stereo-microscope; the presence
of fungi, if any, is observed and recorded. Fungi affects the physical appearance of the seeds.

2. Blotter method: Seeds are typically surface sterilized with a dilute hypochlorite solution and planted on blotters.
These are incubated and observed for 7-10 days. Fungal growth is recorded and confirmed with microscopic
examination. This method typically used for Fusarium, Alternaria, Cercospora (Frogeye).

3. Culture Plate method: Seeds are typically surface sterilized with a dilute hypochlorite solution and placed in 10mm
culture plates with growth media. These are incubated and observed for 7-10 days. Fungal and/or bacterial growth is
recorded and confirmed with microscopic examination. Typically used for Ascochyta, Cercospora (purple blotch),
Botryiscineria, Alternaria.

4. ELISA (enzyme-linked immunosorbent assay): Seeds are typically ground then extracted with a standard extraction
buffer. The solution is then collected and analyzed using 96 well plated coated with specific antibodies. The plate is read
on an ELISA plate reader alongside positive and negative controls. Commonly used for viral infections like Soybean
mosaic virus, bean pod mottle, other viruses.

5. Wash test: Seeds are washed for an extended period and resulting solution is observed under a microscope and any
present pathogens are recorded. Commonly applied for Loose Smut, Soybean Cyst Nematode.

6. PCR (Polymerase chain reaction): Seeds are extracted for contaminating pathogen DNA (Molecular biology
methods). Solution is examined by PCR alongside positive and negative controls. Activity: examine different seed
samples and identify the possible diseases in them. Compared the diseased seeds with reference sample maintained in
the seed disease library to confirm. Applied for Bacterial Fruit Blotch.

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 : SEED ENHANCEMENT
6.1. Introduction
Seeds have evolved over time to respond to a variety of environments. As a result, these adaptations generally produce
satisfactory performance in a range of environments. Seed enhancement technology has a central objective to further
improve seed performance under very specific region and with certain planting equipment. Various techniques have
been employed to assure this superior performance and most have found commercial application.

Quality of seed can be deteriorated due to many reasons as environmental conditions not favourable at the time of seed
formation, mishandling during harvesting, processing and storage and unsuitable storage conditions with high moisture
and temperature which increases seed ageing. The quality of the seed can be improved by seed enhancement technique.
Seed enhancement improves hygiene and mechanical properties, breaking of dormancy, synchronize germination, apply
of nutrients and impart stress tolerance. One such technique is seed priming which could improve seed germination and
germination synchrony in plants. Seed priming is the physiological enhancement technique by which controlling the
hydration level within the seeds so that the metabolic activity necessary for germination could occur but radicle
emergence is prevented.

Although seed quality is governed by genetic make-up; the quality of seeds may deteriorate in subsequent stages like
harvesting, threshing, processing and storage period. Retention of seed germination always important in agricultural
practices. Poor seed handling condition gives rise to deterioration of seed quality and results in the loss of viability.
Also, this greatly affects seed vigour, which ultimately gives poor performance in field and the seed is not able to meet
the quality standards prescribed for that crop.

The productions and timely supply of quality seeds to the farmers are most crucial and challenges the technology. Good
quality seed acts as a catalyst for realizing the potential of all other inputs in agriculture. Without good seed, the
investment on fertilizers, water, pesticides and other inputs will not pay the desired dividends. Therefore, production of
quality seed and maintenance of high germination is utmost significance in the seed program. In this way, seed
enhancements technology has played a significant role in improvising the seed performance.

Seed quality enhancement defined as post-harvest treatment that improve germination or seedling growth or facilitate
the delivery of seeds and other materials required at the time of sowing.

Objectives of seed quality enhancement technology:

✓ Reduce seed rate.
✓ Early emergence and reduce time of emergence under stress conditions.
✓ Supply of growth regulators/ nutrients/ beneficial microbes.
✓ Better nursery management.
✓ Helps seedling to dominate weeds in competition for nutrition.
✓ Field stands and uniformity.
✓ Minimum exposure to toxicant.
✓ Direct seeding of conventionally transplanted vegetable seeds.
✓ High turnover.

Classification of Seed Enhancement Techniques

A. Physiological techniques
Seed hydration/hardening:

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 ✓ Pre-hydration a). Seed fortification b). seed infusion
✓ Priming a). Osmo priming b). Halo priming c). Bio priming c). Hydro priming d). Solid Matrix priming or matri
conditioning
B. Physical techniques
✓ Seed coating (seed coloring, film coating, pelleting and encrusting)
✓ Nano technology (Nano priming)

6.2. Seed Enhancement Techniques Classification

6.2.1. Seed hydration technology (Physiological techniques)
Seed hydration is a process whereby seeds are hydrated using various procedures and then re-dried to permit routine
handling. The objective of seed hydration technology is to increase the percentage and rate of germination, expand the
range of temperatures over which the seed will germinate, and increase the uniformity of stand establishment. To
accomplish these objectives, seeds must be hydrated in some way at a moisture level sufficient to initiate the early events
of germination (Phase I of imbibition) but not sufficient to permit radicle emergence & growth (Phase III).

I. Pre-hydration: A method in which water is freely available and not restricted by the environment. Pre-hydration is
the process of soaking seeds in water or dilute solution of growth regulating compounds to induce early germination,
better root growth, seedling growth and also enhances the yield potential of the crop variety.

There are two types of pre-hydration treatment:

A. Seed fortification: It is pre hydration technique where seeds are soaked either in water or dilute solution of bioactive
chemicals such as micro nutrients, growth regulators, vitamins and seed protectants.

B. Seed infusion: It is a method of impregnation of seeds with bioactive chemicals through organic solvents instead of
water this technique of infusion which helps to avoid the damage caused to the seed due to soaking in water. Hence this
method is highly suitable to the seeds that suffer from soaking or seed coat injury (pulses).

II. Priming: It is based on the principle of controlled imbibition, to a level that permits pre germination metabolism to
proceed, but prevents actual emergence of radical. It is of following types.
A. Hydro priming (drum priming): It is achieved by continuous or successive addition of a limited amount of water
to the seeds. A drum is used for this purpose and the water can also be applied by humid air. 'On-farm steeping' is the
cheap and useful technique that is practiced by incubating seeds (cereals, legumes) for a limited time in warm water.

✓ Treatment usually involves immersion or percolation (up to 30⁰C for several hrs.), followed by draining and drying
back to near original standard moisture content (SMC).
✓ Short ‘hot-water steeps’ (thermotherapy), typically 50⁰C for 10 to 30 min, are used to disinfect or eradicate certain
seed borne pathogens; extreme care, precision needed to avoid loss of seed quality.
B. Halo priming: Halo priming involves the use of salts of chlorides, sulphates, nitrates etc. This priming makes seeds
to improve their performance under salt stressed conditions.
C. Bio priming: It is a process of biological seed treatment that refers to combination of seed hydration (physiological
aspect of disease control) and inoculation (biological aspect of disease control) of seed with beneficial organism to
protect seed with the help of beneficial fungi and bacteria.

Biological Seed Treatment/Bio priming

✓ One of the best alternates to chemical method.
✓ Uses various Biocontrol agents.
✓ Provides protection to seed coat of antagonists (Commonly protect from fungal and bacterial antagonists)

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 ✓ Safe for environment and human health.

Bio-priming is nothing, it is just….

Hydration
of seeds
Bio-agents Bio-priming of seeds

Layer of biocontrol agent is applied over the seed surface as protective coating.

D. Osmo priming or osmo conditioning: is the standard priming technique. Seeds are incubated in well aerated
solutions with a low water potential, and afterwards washes and dried. The low water potential of the solutions can be
achieved by adding osmotica like mannitol, polyethylene glycol (PEG) etc.
E. Solid matrix priming or matri conditioning: It is the incubation of seeds in a solid, insoluble matrix with a limited
amount of water. This method confers a slow imbibition. Matric carriers are Calcinated clay, Vermiculite, Peat Moss,
Sand, Micro-Gel, etc.

6.2.2. Physical techniques

I. Seed Coating: Application of coating substance to the seed to enhance seed placement and performance without
altering shape or placing chemicals on the seed coat which regulate and improve germination.

Seed coating:
✓ Enables accurate and even dose of chemicals and reduces chemical wastage.
✓ Improve the appearance and dust free handling.
✓ To apply fungicides, insecticides, micronutrients directly to seed.
✓ Allow easy flow of seed in automatic seeding.
✓ Act as a temperature switch and water intake regulator.
Steps in Seed Coating Techniques
✓ Seed polymer coating: Polymer + Active ingredients (F+I) Polymer coated seed
✓ Seed coloring: Natural/synthetic dyes Colored seed
✓ Seed pelleting: Adhesives Filler material Active ingredients Pelleted seed

Types of Coatings
A. Film coatings: It’s a sophisticated process of applying precise amount of active ingredients in the form of thin film
along with the liquid material directly on to the seed surface without obscuring its shape.
B. Seed pelleting: It is the process of enclosing a seed with a small quantity of inert material just large enough to
facilitate precision planting or it is the mechanism of applying needed materials in such a way that they affect the seed
or soil at the seed soil interference.

Advantages of seed pelleting: Increase seed size, attraction of moisture, supply of growth regulators, nutrients, saves
chemicals/fertilizers application, uniform field establishment and Increase yield.
Why inert materials are used for seed pelleting? It creates natural water holding media and provide small amount of
nutrients to younger seedlings.

II. Nano Priming: Nano particles (microscopic particle with at least one dimension less than 100 nm) used for priming
with an object to increasing, germination percentage, seedling growth and seedling vigour. Nano priming enhances
germination percentage, seedling dry weight, seedling vigour at 0.02% Titanium dioxide (TiO2) solution in green gram.

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 : GENERAL PRINCIPLES OF SEED PRODUCTION AND MAINTENANCE
Production of good quality seed is an exacting task requires high technical skills and heavy financial investments. During
seed production strict attention must be given to maintain genetic purity and exploit its potentiality in next generation.
In other words, seed production must be carried out under standardized and well-organized condition.

7.1. Genetic Principles of Seed Production

7.1.1. Variety evaluation and development
The breeder initially evaluates new material at the breeding station. Such trials are often not sufficient and more
experimental sites, located it, different ecological zones, are required to obtain a reliable agronomic value for
experimental varieties. In comprehensive seed programs, the final evaluation is usually carried out by a separate varietal
evaluation agency. Varieties from different breeders are objectively compared with existing varieties at a large number
of locations with a wide range of soils and climatic conditions.

A variety submitted by a breeder is not ready for multiplication and marketing until it has undergone extensive variety
research. A farmer simply cannot carry out extensive comparative trials of many varieties; only government institutes,
with trial farms throughout the country, can perform such evaluations to suit the various conditions of different farms
(i.e., varieties appropriate for mechanized operations, different edaphic and climatological conditions, and different
sowing and harvest times). The proposed variety must also be established as an actual new variety. It must be subjected
to botanical variety research for distinguishing characters; at least one character should be found to establish that it is a
new. Based on distinguishability or identity, uniformity, and stability, the Board for Plant Breeder's Rights then registers
the variety in the "Register of Varieties." For agricultural crops, the agricultural value must also be established, and must
be better than that of already existing varieties in order to appear on the Variety List. This list is "binding" for agricultural
crops, which means that only varieties on the list can be used commercially (with some exceptions for grasses and
clovers, and for limited quantities of new varieties still undergoing varietal research; i.e., small-scale trials in practice).

7.1.2. Mechanisms of seed deterioration and maintenance

The genetic purity of a variety or trueness to its type deteriorates due to several factors during the seed production cycles.
The following important factors responsible for deterioration of varieties:
1. Developmental variations
2. Mechanical mixtures
3. Mutations
4. Natural crossing
5. Minor genetic variations
6. Selected influence of pest and diseases
7. The technique of the plant breeder

Genetic purity of seed can be maintained by reducing factors that cause deterioration.
S.
Varietal
The reason for varietal deterioration How to maintain genetic purity
Deterioration
No.
1. Developmental Grown in different environments, soil, climate Growing seeds in their area of
variation and photoperiods. It may set in as differential natural adaptation and growing
growth responses. season to minimize developmental
shifts.

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 2. Mechanical Seed mixture at sowing time, harvesting and - Rogue the seed fields critically
mixture (The threshing time. If presence of volunteer plants - Care should be given during seed
most common) and different varieties grown in adjacent fields. production and processing
3. Mutation Can’t be detected, not so serious. It is often - Remove mutant plant by rouging
difficult to identify or detect minor mutations from seed plots
occurring naturally.
4. Natural - The deterioration sets in due to natural Maintenance of isolation distance is
Crossing crossing with undesirable types, diseased the most important factor in
plants, or off types in cross pollinated crops. avoiding contamination of the cross-
- In self-pollinated crops, natural crossing is pollinated crops.
not a serious source of contamination unless
variety is male sterile and is grown in close
proximity with other varieties.
5. Minor Genetic It often appears in cross pollinated crops. The Care should be given during
Variation variations may lose during later production maintenance of nucleus and breeders
cycles due to selective elimination by the seed of cross-pollinated varieties of
nature. crop is necessary.
6. Selective - New crop varieties often are susceptible to Production of disease-free stocks
Influence of newer races of pests and diseases.
Diseases - The vegetatively propagated stock also can
deteriorate quickly if infected by virus, fungi
or bacteria.
7. The - Premature release of variety - Careful handling and adaptation of
Techniques of - Heritable variation due to recombination's techniques by breeders
the Plant and polyploidization -Periodic testing of varieties for
Breeder - Environmental conditions genetic purity.

7.2. Agronomic principles of seed production

Standardized seed production, besides genetic principles, involves the application of the following agronomic principles
to preserve good seed quality and abundant seed yields.

▪ Selection of suitable agro-climatic condition: The seed crops have to be grown only in areas well adapted to the
photoperiodic and temperature conditions prevailing.
▪ Selection of seed field and its preparation: Seed production field should have good texture and fertility. Should
be free from volunteer plants weeds and other crop plants and also free from soil borne diseases and insect pests.
The land must be prepared well. Good land preparation helps in improved & uniform germination resulted in good
stand establishment.
▪ Selection of variety: Variety should be adopted to agro-climatic condition, high yielder and posses’ other desirable
characters like disease resistance, earliness, grain quality etc.
▪ Isolation of seed crops: The seed crop must be isolated from other nearby fields from the same crop or any
contaminating crop as per certification standards. Time isolation could also be used in some crops. This is a must to
meet the standards for genetic purity.
▪ Seed treatment: If the seed is not treated already, it should be treated with appropriate fungicides/ insecticide prior
to sowing.
▪ Time and method of planting: Should be sown at their normal planting time and sown in rows.
▪ Nutrition of the seed crop: Important role for proper development of plant and seeds.
▪ Seed rate: lower seed rates for easy rouging.
▪ Depth of sowing: small seeds sown upper, larger seeds at more depth.

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▪ Weed control: clean seedbed, clean weed, use herbicide if required.
▪ Irrigation of seed crop: required for translocation of all the nutrients. Seed production areas should be dry regions
with assured irrigation.
▪ Supplementary pollination: rearing honey bees’ aid.
▪ Field inspection: one at flowering and another on grower request.
▪ Rouging: refers to the act of identifying and removing plants with undesirable characteristics from seed fields. rogue
at vegetative, flowering and maturity stage.
▪ Plant protection: disease and insect control.
▪ Harvesting of seed crop: fully matured seed.
▪ Drying of seeds: dry seeds up to 9-12 %.
▪ Storage of seed: less than 21oC.
▪ Seed packaging: seed packed in cotton, jute bag or plastic bag.

7.3. Biotic Stresses in Seed Production

Weeds: objectionable in all crops. Reason
▪ They compete for soil water, nutrients, light etc.
▪ Smolder the crop in delay harvesting and ripening
▪ Impedes cultivation
▪ They are poisonous
▪ They are parasites which weed, broom rape
▪ Harbor pests and diseases.
In seed crop, they are containment if harvested with the crop seeds. There are standard methods of weed control e.g.,
rotation of crops, drainage, flooding, apply fertilizer to promote completive crop growth, herbicides, destroy weed plants.

Diseases and pests: To a certain extent, the incidence of diseases and pests in a crop is influenced by climate as well as
by their presence in the soil. One has to take this into consideration in selecting farms for seed multiplication. To control
the diseases and pests, use the same basic control measures as for food and forage crops e.g., burial of plot debris by
ploughing, rotation of crops, seed treat and insecticidal sprays, isolation of farms from area of with incidence of air
borne and insect borne diseases, (1km distance), good sanitation in seed stores, roguing, special prevention against
rodents and birds.

7.4. Seed production in self- and cross-pollinated crops

A. Self-pollinated crops
Generally, self-pollinating crops have very low outcrossing percentage and using recommended isolation distances will
reduce outcrossing to a great extent. For varieties of such self-pollinating species the following techniques can be used
to produce breeder seed:
▪ Produce enough breeder seed of the variety (need sophisticated storage facility)
▪ Produce breeder seed every fifth year
▪ Use basic seed as source for breeder seed without selection
▪ Use basic seed as source for breeder seed with negative mass selection
▪ Ear- to- row or plant-to- row selection (pure-line selection)

The best approach for breeder seed maintenance and production of self-pollinated crops is an ear-to- row or plant-to-
row procedure. Single ear or single plants typical of the variety are selected and harvested separately. The seed of each
ear or plant are carefully observed, and any (or plant) that produces one or more deviant seeds is discarded. The seeds

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of selected ears (or plants) are planted in ear rows (or plant rows). The ear rows are periodically examined throughout
the growing season for wheat, at least four inspections should be made: i) early growth stage; ii) before ear emergence;
iii) at ear emergence and iv) at maturity. The varietal characteristics are clearly expressed at these stages. Selected ear
rows are bulked up to constitute varietally pure and disease- free breeder seed, which is used to initiate the seed
production cycle (pre-basic seed, basic seed and certified seed).

Variety maintenance scheme for self-pollinated crops

Select 400-500 ears of wheat at harvest time on the basis of complete uniformity

Thresh each ear separately

Sow the seed from each ear in a single labeled row, normally of 20-25 plants (= an ear row)

Inspect each ear row very carefully and discard any rows in which there is an off-type

At harvest select a further 400 heads of ideal type to provide ears for the following year. Bulk the remaining seed to
form a breeder seed lot.

B. Cross pollinated Crops

1. Maintenance and production of breeders’ seed of cross-pollinated crops
The maintenance of varietal purity of cross-pollinated crops, such as open pollinated varieties of maize and sorghum,
may be done: Either in isolated fields or under controlled pollinations. The plot used for breeders’ seed should not be in
a field that had the same crop in the previous season.

A high standard of management is also required to ensure that the crop is grown under optimum conditions. During
vegetative growth, off-type and variant plants should be removed, so that, at flowering, they do not contaminate the true
variety. The maintenance and production of breeders’ seed of a cross pollinated crop variety depends on whether the
variety is an open-pollinated or hybrid variety.

2. Production of breeders’ seed of open-pollinated maize varieties in open-pollinated fields

Where the breeders’ seed is produced in open-pollinated fields, it is essential that the crop is sufficiently isolated from
potential contaminant crops to ensure that the breeders’ seed is pure.

Two methods of isolation may be employed.

▪ Isolating by distance from a contaminant crop.
▪ Isolating by time, in which case the breeders’ seed plot is planted in close proximity too, but by a time gap sufficient
to avoid overlap of the flowering periods. For maize, a time gap between plantings of the seed plot and the
contaminant crop must be more than 28 days.

The plot may be laid out in two ways:

A. Male and female rows may be identified, with the female plants detasseled prior to flowering to ensure cross-
pollination from the male rows. The seed from the female rows is carefully selected from typical plants and desirable
ears, with seed from at least 400 plants bulked for breeders’ seed. The seed from the male rows may be discarded or
used as basic seed.

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B. The plot is grown as an open-pollinated field, with no separation into male and female rows. At least 600 plants that
are typical of the variety are selected from within the field, and the seed from the central portion of the cobs is bulked
for breeders’ seed.

7.5. Hybrid seed production

In order to better appreciate the methods used for the seed production, it is essential to understand the pollination
mechanisms particularly in cross pollinated crops. Hybrid is produced by crossing between two genetically dissimilar
parents. Pollen from male parent (pollen parent) will pollinate, fertilize and set seeds in female (seed parent) to produce
F1 hybrid seeds. For production of a hybrid crossing between two parents is important, the crossing process will result
in heterosis. In self-pollinated cross it is difficult to cross but in cross pollinated crops it is easier.

In nature to create genetic variability and for its wider adaptation in different environmental conditions, flowering plants
has adopted many mechanisms for cross pollination. Cross-pollination results in genetic heterogeneity and show wider
adaptations. Flowering plants have evolved a number of devises to encourage cross-pollination. Those mechanisms are;

A. Dicliny: Flowers are unisexual. In monoecious plants male and female flowers are borne on the same plant e.g.,
maize. In dioecious plants male flowers are borne on different plants. e.g., papaya.

B. Dichogamy: Stamens and pistils of hermaphrodite flowers may mature at different times facilitating cross –
pollination. The time gap between the two may vary from one day to many days.
▪ In protandry anthers dehisce earlier than the stigma receptivity. e.g., maize.
▪ In protogyny stigma become receptive earlier than the anthers dehisce. e.g., Pearl millet.

C. Self-incompatibility: Self-fertilization is avoided by recognizing the self-pollen by the stigma. Such flowers unable
to be fertilized by its own pollen. e.g., Cole crops/Brassica.

D. Herkogamy: There is spatial separation of the anthers and stigma. The irrelative position is such that self-fertilization
cannot occur. The stigma projects beyond the anthers and therefore pollen cannot land on stigma. E.g., Lucerne stigma
is covered with a waxy film. The stigma does not become receptive until this waxy membrane is broken by visit of
honey bees resulting cross pollination.

E. Heterostyly: In such flowers the stigma and anthers grow at different heights which does not favor self-pollination.
The condition of having unequal styles. e.g., Brassica

F. Male sterility: Absence or atrophy or mis or malformed of male sex organ in normal bisexual flower (inability to
produce viable pollen). e.g., Sunflower. Types of Male sterility: Genetic male sterility, Cytoplasm sterility and
Cytoplasmic- genetic male sterility.
G. A combination of two or more of the above mechanisms may occur in some species. This improves the efficiency
of the system in promoting cross pollination

Characteristics of parental lines

Female Parent Male Parent
High seed yield Good pollen production
Good seed characteristics Long shedding period
Male sterility Plant height
Lodging resistant Fertility restoration

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Basic procedures for hybrid seed production
A. Development and identification of parental lines
B. Multiplication of parental lines
C. Crossing between parental lines and production of F1

Depending on the type of progenitors, hybrid cultivars may be classified as:

1. Single cross: Hybrid seed produced by controlled crossing between two selected inbreds (A×B).
2. Double cross: Hybrid produced by crossing between two certified single crosses (A×B) × (C×D).
3.Three way cross: Hybrid seed produced by crossing between an inbred used as male and certified single cross hybrid
as female parent [(A×B) × C].
4. Top cross: hybrid seed produced by crossing of inbred line with a certified open pollinated variety.
5. Double top cross: Hybrid seed produced by crossing between a certified single cross and a certified open pollinated
variety.

Designation of classes of Seed in Ethiopia

The new varieties, released by private or government must pass through a series of evaluation, release and registration
tests and procedures before farmers can use them. Seed production of registered cultivars follows a generation system
to ensure that all seeds marketed to farmers should be originated from a known source (breeder seed). Each generation
is produced under strict supervision and must meet seed quality standards. Ethiopia has adopted from Organization for
Economic Cooperation and Development (OECD) seed generation scheme. The four generally recognized classes of
seeds are: Breeder seed, pre-basic seed, Basic seed and certified seed.

1. Breeder seed (1st generation) is the initial source of seed and is usually produced by the breeder. It is the source for
the production of pre-basic or basic seed. 1st generation supplied by plant breeders.
2. Pre-basic seed (2nd generation) is the progeny of the breeder seed and is usually produced under the supervision of
a breeder or his designated agency. This generation is commonly used for crops that have low multiplication ratios
and where large quantities of certified seed are required.
3. Basic seed (3rd generation) is the progeny of breeder or pre-basic seed and is usually produced under the supervision
of a breeder or his designated agency and under the control of a seed quality control agency.
4. Certified seed (4th generation and above) is the progeny of basic seed and is produced on contract with selected
seed growers under the supervision of the seed enterprise, public or private. Certified seed can be used to produce
further generations of certified seed (certified 1, certified 2, certified 3, certified 4 etc.) or can be planted by farmers
for grain production.

The number of generations used in the certification system is fixed at the lowest possible level because the genetic value
of a seed crop can, in principle, only decrease from one generation to another. Restricting the number of generations is
a way to preserve quality in cross-pollinating crops this is more important than in self-pollinating crops.

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 : SEED QUALITY CONTROL
8.1. Definitions and Objectives of Seed Quality Control
Definitions:
✓ Quality: - a measure of excellence of an item or product with reference to standards.
✓ Control: - imposing limitation or restriction with reference to standards.
✓ Standard: - reference frame for comparison. It indicates the minimum limits of seed quality attributes.
✓ Seed quality control: - means the process of evaluating the quality of a seed for compliance with a given country
seed standards. e.g., Ethiopian seed standards.

Objectives of seed quality control

➢ To determine approval or rejection of a seed crop or seed lot in order to ensure quality
➢ To offer protection to producers and consumers alike against any kind of fraud and malpractices.

What is quality seed?

Quality seed is genetically pure, characterized by a high germination percentage and appropriate moisture content; it is
free from diseases, and has a high content of pure seeds and no weed seeds. High quality seed is expected to produce
normal seedlings that will emerge well in the field, give the farmer a uniform crop stand and produce a high yield.

What is seed quality: - a concept that expresses the extent to which a given seed lot meets the standards set for certain
attributes determining the quality status of seeds. Such attributes are varietal and physical purity, physiological quality,
seed health and moisture content.

Seed quality control is a multiple concept made up of a number of attributes which comprise:
1. Varietal (genetic) purity: Extent/degree of purity of the variety expressed as a percentage (True to type)

2. Physical (analytic) purity: Indicates how much of the material in a seed lot is pure seed of the species named on the
label. Pure seed is weighed and expressed as percentage by weight of whole sample.

3. Physiological Quality
A. Germination capacity
Germination capacity of a seed lot is percentage by number of pure seeds which produce normal seedlings in a laboratory
test. Weak and abnormal seedlings in any way are ignored. It indicates the potential of a seed lot to establish seedlings
under good field conditions:
▪ Most field conditions are not optimal
▪ Seed lots with higher germination capacity will always prove to establish more seedlings than those with lower
germination capacity, especially under sub-optimal conditions.
Both seed physical purity and seed germination have a profound effect on yield and determine the planting value of the
seed. Planting value (percentage of pure live seed) defines the true value of a seed lot for crop cultivation. Only pure
live seed produces plants, and it should therefore be calculated in order to accurately adjust the seeding rate, as necessary.
Pure live seed (%) = Pure seed (%) x Germination (%)

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Example: If a seed bag tag indicates germination of 80% and purity of 95%, Pure live seed = (80 x 95) / 100 = 76%
Therefore, the seed lot contains 76 kg actual live seed for every 100 kg.

B. Seed vigour
Refers to the ability (“degree of aliveness”) of seed to germinate and continue growth under adverse or sub-optimal field
conditions. Seed lots of apparently equal quality as indicated by germination % will produce different responses in field
emergence.
C. Moisture content
Seeds should retain their germination capacity at the highest possible level during storage to ensure growth into normal
and healthy seedlings. Moisture content and storage temperature are the two factors which have the greatest effect on
viability of stored seeds. The two factors are important because the influence respiration rate not only of the seeds but
also of the fungi and other micro-organisms. Safe moisture contents for safe storage of seeds vary with the type of crops:
12-13% for cereals, 8-10% for legumes and vegetable seeds, 7-8% for oil crops. Moisture content of seed is measured
by moisture meters and by controlled oven dry method.
D. Seed health
In certain crops, seed health is an important factor in the control of crop diseases and field establishment. Seeds may
carry pathogens like bacteria, fungi, viruses and nematodes affecting crops adversely.

8.2. Seed Certification

Seed certification is a regulatory process designed to maintain and make available to farmers high quality seeds and
propagating materials of superior crop varieties, grown and distributed to ensure genetic identity and genetic purity. It
also ensures other factors, including absence of weeds and diseases, analytical purity and viability. The seed
certification authorities adopt predetermined standards and systems for the production, multiplication and marketing
of seed.

Seed certification programs have been in existence for over 100 years. They have effectively defined and monitored
standards to guarantee specific purity requirements of the final product or seed.

Certification means that you can buy seed with high physical and physiological quality standards, as close as possible
to the genetic make-up of the variety selected by the breeder. The plant breeder invests time and effort in selecting a
variety that performs better than the varieties available on the market. Farmers who choose to grow the certified variety
have the benefits of quality seed. For example, in the United Kingdom, there was a 33% increase in national average
wheat yields from 6.2 tons/ha in 1982 to 8.3 tons/ha in 2008 around 90% of which is due to plant breeding. Farmers can
optimize this potential by buying seed of modern varieties with high varietal purity through a certification scheme.

Objectives of seed certification are:

▪ To establish and maintain reasonable standards of seed quality.
▪ To facilitate the production of seed of specified quality.
▪ To maintain the identity and purity of varieties.

I. Seed Certification Process

It is important to maintain varietal purity and seed quality throughout the certification process, and the procedures in
place aim to give continuity. There are four important stages in the certification process:

▪ Control seed in previous generations.

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▪ Carry out field inspections during the multiplication process to ensure there is minimal contamination and that the
variety is true to type.
▪ Test seed quality in laboratories.
▪ Grow samples in control plots of the known seed to ensure that the progeny conform to the characteristics of the
variety.

Seed certification agency

The seed certification agency is the competent authority independent from the industry responsible for implementing
the certification scheme. The organization and the structure of a country’s certifying authority may vary depending upon
the development of the seed sector. In some countries, certification is under public authority and the agency is also
responsible for the seed testing laboratory.

Main activities of a seed certification agency:

▪ Registration of varieties and maintenance of the variety list for seed certification
▪ Establishment and review of seed certification standards
▪ Registration of seed growers
▪ Registration of processing plants for seed certification
▪ Field inspection
▪ Seed sampling
▪ Issue of official labels
▪ Laboratory testing
▪ Awarding of certificates
▪ Seed certification and seed testing training.

Seed Certification Agency should function on the following broad principles:

▪ It should not involve in seed production and marketing
▪ It should be an autonomous body
▪ Seed certification procedure adopted should be uniform throughout the country
▪ It should closely associate with technical and other related institutes
▪ It should operate on a no profit and no loss basis
▪ It should have adequate technical staff and facilities for timely inspection of seed fields
▪ It should serve the interests of seed producers and buyers/farmers

Responsibilities for seed quality control and assurance

The regional authorities shall have the responsibility for quality control and certification of regionally produced pre
basic, basic and certified seed to be released on the domestic market as “Approved Seed” or “Quality Declared Seed”.
The Ministry of Agriculture shall be responsible for the quality control and certification of imported registered seed for
release on the domestic market as Approved Seed, domestically produced seed for export and emergency seed. The
quality control of seeds shall be undertaken in conformity with Ethiopian seed standards.

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Eligibility for certificate of seed quality
In order to be eligible for certificate of seed quality, the seed shall be:
▪ A registered variety.
▪ Produce by a producer who holds valid certificate of competence.
▪ Seed must be obtained from known source.

The issuance of certificate of approved seed shall undergo the following steps:
▪ Field inspection of the seed
▪ Seed processing and sampling
▪ Testing in accredited laboratory
▪ Affixing of tag and seal on packing or containers
▪ Certification fee and other service charges.
▪ Grant of certificate.

Seed certification standards

Seed certification requirements vary according to local conditions and national laws and regulations. The aim is to
provide the buyer with the best possible assurance of obtaining good quality seed of known purity and origin.

Technical seed certification requirements include field (production) standards and seed standards:
• Varietal purity, isolation, seed-borne diseases and weeds: - controlled by field inspections and pre- and post-
control tests.
• Analytical purity, germination, seed heath, vigour and moisture content and variety purity (in so far as possible): -
controlled by seed quality testing.

II. Field inspection

Field inspection means an official checkup of a seed field to check compliance with field standards set by seed agency.
Field inspection standards should contain the following information:
▪ Standard of seed planted
▪ Crop rotation
▪ Isolation distance
▪ Maximum percentage of other varieties or off-types
▪ Maximum percentage of seed-borne disease
▪ Maximum percentage of objectionable weed plants
▪ Minimum number of field inspections required.

Objective of field inspection

To verify the factors which can cause irreversible damage to the genetic purity or seed health.
The most important functions of field inspection are:

▪ To check varietal identity.

▪ To ensure varietal purity.

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Crop inspection procedure
▪ Confirm the crop entry details and the correct location of the field.
▪ Authenticate the seed sown to produce the crop.
▪ Positively identify the variety as far as possible in the field.
▪ Collect information on the cropping history of the seed field and verify whether the seed field meets the
prescribed land requirements.
▪ Detect and record any admixtures of other varieties of the same species.
▪ Assess noxious weed contamination.
▪ Check the isolation requirements.
▪ Assess the general condition of the crop including the amount of lodging and poor growth.
▪ Assess other species contamination.
▪ Check freedom from seed-borne diseases.

Crop stages and number of field inspections

Field inspection has to be done at different crop stages namely, pre-flowering, flowering, post flowering and harvesting
stages. The number of field inspections and the stages of crop growth at which the field inspections should be conducted
will be based on the nature of pollination and type of seed produced (hybrid / population varieties).

Stages of field inspection for different seed propagated crops

Vegetative Flowering Post
Crop Pre harvest
stage stage flowering
Self-pollinated crops - ✓ - ✓
Cross pollinated crops ✓ ✓ - ✓
Hybrids ✓ ✓ ✓ ✓

The field inspection during flowering stage should be made without any prior notice to the seed grower in the seed
production fields of cross-pollinated crops and self-pollinated crops. However, in self-pollinated crops, the seed grower
may be informed about the date of inspection.

Completing the inspection report

At the end of the inspection, the inspector records the number of off-type plants observed and the number of plants per
hectare. With reference to the reject number tables, the number counted is compared with the number permitted. A
recommendation can then be made by the certifying authority. It is the responsibility of the certifying authority to issue
a result for each crop usually not until all field inspections have been completed, but before harvest. The final result is
based not only on the field inspection findings but also on the control plot results; it is important to recognize that the
final result may not be that of the field inspection.

8.3. Seed Legislation

Seed legislation is a regulatory mechanism put in place to protect the farmer from purchasing seed of doubtful quality.
In general, the objective of seed legislation is to regulate seed commerce including variety certification and variety
protection.

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Legislation Support Procedures
Seed Legislation is enacted in two phases
1. Seed Proclamation by Parliament
2. Seed Rules, Regulations and Procedures by Minister of agriculture

The Seed Proclamation

A document enacted by parliament which lays down the general principles that the seed act is to declare, purpose of
seed legislation and how to achieve it. A seed law must be enacted to protect the farming community against fraud,
negligence or accident.

Variety and Seed Rules and Regulations

▪ Detailed procedures for enforcing the seed law
▪ After enacting the seed proclamation, the Minister of Agriculture is empowered to formulate appropriate rules and
regulations.
▪ They are put into effect and can be amended whenever necessary, to keep pace with the development of the seed
program, without taking valuable parliamentary time.

Seed Law Enforcement by Certification Agency (CA)

▪ After a Seed Law is enacted; certifying authorities are designated and given responsibility and legal powers for
implementing the Seed Act.
▪ Inspectors of the CA must be well facilitated to carry out Seed Law enforcement in order to be effective.
▪ Training of inspectors essential to enforce the law.

Seed Legislations in Ethiopia

The formal seed quality system was partially exercised in a much-disorganized manner in some parts of the country as
early as 1942. There was no organized system in the country until the government the establishment of the Ethiopian
Seed Corporation (ESC) in 1987, which is state owned. The ESC was the only state-owned seed institution charged with
seed production and marketing in the country until 1990. The Ethiopian seed policy was first issued in October 1992,
and serves as the basis for different laws and regulations. The 1992 seed policy was replaced by Proclamation No.
56/1993 for the establishment of the National Seed Industry Agency (NSIA). Seed Regulation No. 16/1997 was issued
by the Council of Ministers. The seed system was legally enforced in 2000 through the seed law No.206/2000.
Eventually, the 2000 seed law replaced by still functioning seed law Proclamation No. 782/2013.

Recently, the Plant Breeders’ Rights proclamation and the Access and Benefit sharing Law were issued, however these
policies still need to be put into practice, through the establishment of regulatory frameworks. Despite the existence of
a seed policy, seed law and seed standards, their implementation is still mostly at the infant stage. Seed certification and
quality control has been decentralized to the regional seed enterprise that make use of their own seed laboratories.
However, these laboratories and services are limited in terms of equipment, skilled man power, financial and operational
capacity. Furthermore, since seed quality control has been embedded within the Bureaus of Agriculture, they are also
responsible for organizing production and dissemination.

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 : SEED SUPPLY SYSTEM
Seed systems are composed of set of dynamic interaction between seed supply and demand, resulting in farm level
utilization of seed and thus plant genetic resource. The term seed system represents the entire complex organization,
individual and institution associated with the development, multiplication, processing, storage, distribution and
marketing of seed in one country.

The seed system includes traditional (or informal) system and the non-traditional (or formal or commercial) systems.
Legal institutions such as variety release procedures, intellectual property rights, certification programs, seed standards,
contract laws, and law enforcement are also an important component of the seed system of any country. They help to
determine the quantity, quality, and cost of seeds passing through the seed system.

Seed system participants may be relatively few or many, predominantly public or private depending upon the farmers
that the system serves. In local systems of seed exchange, farmers often undertake most of the activities that define a
seed system. As systems expand to national, regional, and international scales, participants will include the following:
farmers, international agricultural research centers, private and public domestic seed enterprises, retailers and
distributors, multinational seed companies, private research institutions, farmers associations and cooperatives, banks
and credit institutions, trade associations, local governing bodies, donor agencies, national agencies and ministries,
community groups (social, religious, etc.), agricultural universities, national agricultural research institutes and
NGOs/PVOs. These participants may assume multiple roles in the process of seed provision, performing one or several
activities.

Seed systems, formal or informal, fulfill a series of functions that are basic prerequisites for expecting the best possible
productivity from a crop in a specific situation. Healthy, viable seed of the preferred variety needs to be available at the
right time, under reasonable conditions, so that farmers can use their land and labor resources with the best yield
expectations.

Seed systems in Ethiopia can be divided into two broad types: the formal system and the informal system (sometimes
called local or farmers seed system). Both systems are operating simultaneously in the country and difficult to demarcate
between the two. However, a fact that the formal system is the original source of improved seeds in the informal system.
There is also a system referred to as integrated seed system. Other forms of seed systems operating in both systems
also exist such as Community Based Seed System (CBSS). However not well developed, few commercial seed
systems, as part of the formal system, are also operating in the country.

9.1. Formal Seed Supply System

The formal seed supply system is highly regulated and involves a chain of activities leading to clear products which are
certified seed of verified varieties. The chain usually starts with plant breeding and selection, resulting in different types
of varieties, including hybrids, and promotes advanced fixed germplasm materials leading to formal variety release and
maintenance. Guiding principles in the formal system ensure that varietal identity and purity are maintained throughout
the various generations of seed multiplication (Breeder, Pre-basic, Basic and Certified seed in some cases Commercial),
with optimal physical, physiological and sanitary quality. Conscious efforts are made to ensure that the appropriate
documented multiplication process and procedures are followed and that quality control measures are taken at various
stages of the operation.

Private seed enterprises (private sector) and public seed sectors are in the domain of the formal seed supply system and
the bulk of seed generated through this system covers economically viable crop species with good recurrent seed

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demands, such as vegetables, hybrids and some cross-pollinated crops (maize for example) and to a lesser extent some
self-pollinated crops (wheat, barley for example) or some vegetative propagated crops (mainly potato).

The system has been referred to by other names including:

a. Organized seed system
b. Conventional seed system
c. Commercial seed system and
d. Regulated seed system.

9.2. Informal Seed Supply System

The informal seed supply system refers to the traditional arrangements used by farmers to supply the seeds they need to
plant in the following season. Other names given to informal seed supply systems include:
a. Farmer-managed seed system
b. Farm-based
c. Local seed production and supply
d. Traditional seed system; and
e. Farmers’ seed system.

Activities tend to be integrated and locally organized and embrace most of the ways in which farmers produce,
disseminate, and access seed: directly from their own harvest; through exchange and barter within the community; and
through local markets. It is a flexible system and varieties of seed may comprise landraces or old or new improved
varieties; however, the seed is of variable quality. The same general steps or processes take place in the informal seed
supply system as in the formal sector (variety choice, variety testing, introduction, seed multiplication, selection,
dissemination and storage) but they take place as an integral part of farmers’ production systems rather than as discrete
activities. While some farmers treat “seed” as special, there is not always necessarily a distinction between “seed” and
“grain”. The steps do not flow in a linear sequence and are not monitored or controlled by government policies and
regulations. Instead, they are guided by local technical knowledge and standards and by local social structures and
norms.

The relative importance of these two systems varies depending on the state of development of the agricultural system
and the crops. About 80 percent of food production reportedly comes from farmers with smallholdings and the majority
of farmers in developing countries use seed from the informal seed system. Most of the seed covered by this system falls
within crop groups that are not of commercial interest to the private sector but the bulk of which constitute important
food security crops. Contrary to conventional views, the formal and informal seed delivery systems coexist in large part
in developing countries and in some cases in developed countries; farmers will usually resort to either or both of these
systems for different crops and for different seasons.

According to Cromwell et al. (1992), five key features distinguish the informal from the formal system:
a. Traditional system
b. Semi structured
c. Operate at the individual or community level
d. Uses a wide range of exchange mechanisms, and
e. Usually deal with small quantities of seeds often demanded by farmers.

In Ethiopia, the informal system is extremely important for seed security. The bulk of seed supply is provided through
the informal system, implying its importance in national seed security. About 60-70% of seed used by Ethiopian

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smallholder farmers is saved on-farm and exchanged among farmers, and the remaining 20-30% is borrowed or
purchased locally. The informal seed system (either self-saved seed or farmer-to-farmer seed exchange) accounts for
90% of the seed used by smallholder farmers (Belay, 2004), while the share of improved seed is less than 10%.

The majority of Ethiopian farmers show a tendency of depending on the informal system due to the following key
reasons:
a. It is relatively cheaper and readily available in the farmer’s villages just at the time of seed is needed.
b. It allows use of seeds after testing on primary adopter farmers.
c. It is more reliable and its sustainability is more guaranteed than the formal system.

9.3. Integrated Seed Supply System

The line between the formal and informal seed sectors can become somewhat blurred, as seeds of improved varieties
can be saved by farmers and eventually considered as “local variety” or “local seed” after some years of usage. In
addition, in Ethiopia there have been attempts made by the government and NGOs to promote quality seed production
and distribution through market channels for landrace varieties, although until now the volume they represent is quite
small. Thus, the formal and local seed systems are not always as distinct or separated as the two labels may imply
something to integrate and synergize both systems.

It is obvious that the two systems are interacting in many ways and this interaction is found to be beneficial. Integrating
the formal and local systems is, therefore, important to exploit benefits of the synergetic impacts as a result of integration
on addressing seed security and sustainability in the country.

9.4. Seed Industry Development in Ethiopia

The Ethiopian seed industry is composed of formal and informal sectors as well as public and private organization. The
formal sectors include federal and regional agricultural research establishments, universities, the regulatory organ in the
MoARD, regional seed enterprises and private companies. The informal sectors encompass millions of farmers, who
continue to practice seed selection and preservation, just as their ancestors did.

The formal seed sector in Ethiopia owes its origin to the establishment of Jimma agricultural college in 1942, the
Alemaya College of Agriculture (currently HrU) in the mid-fifties, and the Institute of Agricultural Research (IAR) in
the mid-sixties, when these institutions engaged in developing improved varieties of seeds for cereal, pulse and oilseed
crops. Seeds developed by these institutions initially introduced to the farmers in 1967. This development led to
increased demand for modern agricultural input, particularly improved seed. While the provision of other agriculture
input from local and foreign sources was possible, improved seed supply was lacking as there was no organized system
in the country until the government the establishment of the Ethiopian Seed Corporation (ESC) in 1987, which is state
owned.

Initially, the ESC was established not in view of addressing the seed need of small-scale farmers, but, following the
policy of the then socialist government, which believed to bring food self-sufficiency by producing much on such farms
to supply seed to the State and producer cooperative farms. The situation undermined the larger majority of the private
small-scale farmers who produced the bulk of the grain that the country harvested. The ESC was the only state-owned
seed institution charged with seed production and marketing in the country until 1990. Like in any other sector, by then,
private initiatives were not allowed to enter into crop breeding and seed businesses.

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The ESC therefore, sold the lion’s share of the seed it produced to the State-owned farms, which remained its major
client until the early 1990's. Although its activities were largely skewed to the state farms and cooperatives at the expense
of small farmers, the establishment of the ESE did lead to the start of an organized seed production and supplier in the
formal sector.

Lately important developments have taken place in Ethiopia in respect to government involvement. The decision was to
decentralize the public research infrastructure and the regional government establishment of their own Seed. Then seven
regional agricultural research institutes in the country that were established since 1997 under the administration of
individual regional states to undertake agricultural research specific to the region's agricultural sector. These are the
Tigray Agricultural Research Institute, Amhara Regional Agricultural Research Institute, South Agricultural Research
Institute, Oromia Agricultural Research Institute, Somali Pastoral and agro-pastoral Research Institute, Afar Agricultural
Research Institute, and Gambella Agricultural Research Institute.

The other improvement is establishment regional government seed enterprise aims on the one hand at reducing costs
and to make production of seeds more profitable; on the other hand, it should support the emergence of small and local
seed companies that can more directly aim at satisfying the diverse needs of farmers. In parallel, the changes in terms
of seed quality control and the introduction of the concept of quality declared seed fit very much in the development of
providing more opportunities for the local private sector. Secondly, the proclamation on plant breeder’s rights signifies
a clear message to the commercial sector that it has an important role to play in Ethiopia. It aims at increasing access to
foreign varieties (important for export horticulture) and at stimulating plant breeding at home.

To create the right condition for the establishment of strong seed system for production and supply of good quality seed
to the farming community, the government formulated the national seed industry policy, which was issued in October
1992. The policies are instrumental to developing a healthy national seed industry conserving and sustain genetic
resource, reinforcing crop breeding research and supplying of high-quality seed to the farmers to participate in
germplasm conservation as well as in the seed production and supply system. It also has an objective of creating a
functional and efficient institutional linkage among seed industry participants.

MoARD is the umbrella organization which coordinates the leads the various activates of the seed industry in Ethiopia.
The main tasks of MoARD`s various department include the national seed policy, variety registration and release, seed
import/export, seed certification, quarantine, and extension. Next, the responsibility for the official seed quality control
and certification was given to the National Seed Industry Agency (NSIA) in 1993. Then, it was replaced by the
Agricultural Input Quality Control Department of MoARD.

Currently, the major actors of the formal seed system in Ethiopia are: National Agricultural Research Systems (NARS),
Ministry of Agriculture and Natural Resources (MoANR), Ethiopian Seed Enterprise (ESE) and private seed companies
specializing on specific crops like maize. Recently, regional seed enterprises (RSE) were also established as public seed
enterprises such as; Oromia Seed Enterprise (OSE), Amhara Seed Enterprise (ASE), and Southern Nations
nationalities and Peoples Region Seed Enterprise (SRSE) are entered into the formal seed system. All actors have
inter-dependent roles in the system and inefficiency of one actor will automatically affect negatively the performances
of the rest of the actors. NARS (EIAR & RARIs) is responsible for variety development and supply of initial seed, and
ESE and RSEs are playing key roles in mass production of improved seeds. MoANR is also involved in variety release,
multiplication, certification, and distribution of seeds in the country. Private seed growers and other farmer institutions
such as unions and cooperatives are also playing key roles in multiplication and distribution of different classes of seeds.

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